Cal-t constructs and uses thereof

ABSTRACT

The technology described herein is directed to compositions comprising components of multi-component CALs or CARs, e.g., a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain. Further provided herein are methods for treating or preventing an autoimmune disease, a transplant rejection, or graft versus host disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 62/916,924 filed Oct. 18, 2019, the contents of which are incorporated herein by reference in their entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Oct. 16, 2020, is named 701586-096030USPT_SL.txt and is 540,018 bytes in size.

TECHNICAL FIELD

The technology described herein relates to chimeric antigen ligand (CAL) technology, e.g., for treating of autoimmune and/or T-cell mediated conditions.

BACKGROUND

The major cause of autoimmunity and transplant rejection, e.g., following an organ transplant, is autoreactive and alloreactive T cells attacking the patient's or donor's organ(s), tissue, and/or cells. Current treatments for preventing autoimmunity and transplant rejection involve stringent immunosuppression, which can lead to severe, unwanted side effects including extreme susceptibility to infection, malignant and benign neoplasms, multiple organ and systems failure. In order to prevent autoimmunity and transplant rejection without inducing unwanted side effects, it would be advantageous to delete autoreactive and alloreactive T cells in a patient receiving an organ transplant without needing to additionally administer immunosuppressant medications, or to reduce the doses of the immunosuppressant regime delivered concurrently. This type of therapy, however, has not been successfully demonstrated.

T cells recognize their target cells by using receptors on their cell surface which are called T Cell Receptors (TCRs). TCRs have a recognition portion and a signaling portion. When the recognition portion binds to the natural complexes formed in the body in the presence of a diseased cell, the signaling portion is activated, which leads to the T cell engaging in killing activity or recruiting other immune cells to destroy the diseased cell. The natural complexes are also known as peptide-major histocompatibility complexes (pMHC complexes). CAR-T cell therapy is known and seeks to help T cells recognize autoreactive and alloreactive T cells. This is accomplished by genetically altering a T cell so that it expresses a chimeric antigen receptor (CAR). The CAR is an altered TCR, in which the natural recognition portion is removed and replaced with a synthetic recognition portion that is designed to more effectively recognize the autoreactive and alloreactive T cells by very specifically detecting the presence of a molecule unique to the autoreactive and alloreactive T cells. These CAR-T cells are then given to a patient. Inside the patient, their synthetic CAR molecules will bind to the autoreactive and alloreactive T cells and in the act of that binding, activate the T cell, resulting in the patient's own immune system attacking the diseased T-cells.

SUMMARY

Described herein are methods and compositions relating to chimeric antigen ligands (CAL). The CALs described herein comprise a TCR recognition domain and a biomolecular interaction domain. The TCR recognition permits the CAL to bind to autoreactive and/or alloreactive T cells in an antigen specific manner. The biomolecular interaction domain permits an immune killer cell (e.g., a NK cell, a T cell, or a dendritic cell) to bind to the CAL, thereby promoting killing of the autoreactive and/or alloreactive T cell by the immune killer cell.

The TCR recognition domain of a CAL binds specifically to a TCR, e.g., a TCR expressed on the surface of an autoreactive and/or alloreactive T cell. Exemplary but non-limiting TCR recognition domains include peptide-MHC complexes, e.g., in monomeric, oligomeric, or multimeric form. The TCR recognition domain can comprise natural or synthetic sequences. Specific examples of peptide-MHC complexes that include autoreactive or alloreactive antigens are provided elsewhere herein.

The biomolecular interaction domain of the CAL permits specific binding of the CAL with a second biomolecule, e.g., a receptor on the immune killer cell. In some embodiments, the biomolecular interaction domain of the CAL is recognized by an endogenous receptor on the immune killer cell. In some embodiments, the biomolecular interaction domain of the CAL is recognized by an engineered receptor on the immune killer cell. Exemplary but non-limiting biomolecular interaction domains include FITC (which can be recognized by a FITC CAR-T cell system), a leucine zipper domain, a zinc finger domain, PSD95-D1g1-zo-1 (PDZ) domains, a streptavidin domain and a streptavidin binding protein (SBP) domain, a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1), a PYL domain and an ABI domain, a chemically-induced pair of interaction domains as described elsewhere herein, a Snap-tag, a Halo tag, a T14-3-3-cdeltaC and/or a C-Terminal peptide of PMA2 (CT52), a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein a Fab domain, and/or an anti-CD3 domain.

As noted, the CALs described herein can be used in combination with CAR-T systems. Exemplary but non-limiting CAR-T systems suitable for use with the methods and compositions described herein include SUPRA CAR and SPLIT CAR. CAR-T systems are discussed in more detail elsewhere herein and are known in the art.

In one aspect of any of the embodiments, described herein is a composition, comprising: a) a TCR recognition domain; and one or both of: b) an intracellular signaling domain; and c) a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain).

In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; wherein the first-type and second-type biomolecular interaction domains bind specifically to each other. In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the first-type and third-type biomolecular interaction domains bind specifically to each other. In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; and a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a third-type biomolecular interaction domain; wherein the second-type and third-type biomolecular interaction domains compete for binding to the first-type biomolecular interaction domain. In some embodiments of any of the aspects, the third-type biomolecular interaction domain and first-type biomolecular interaction domain have a higher affinity for each other than the second-type biomolecular interaction domain and first-type biomolecular interaction domain.

In one aspect of any of the embodiments, described herein is a composition comprising: a first polypeptide comprising at least a portion of a TCR recognition domain and a first-type biomolecular interaction domain; a signaling polypeptide comprising a second-type biomolecular interaction domain, a fourth-type biomolecular interaction domain, and an intracellular signaling domain; and a recognition polypeptide comprising a second recognition domain and a fifth-type biomolecular interaction domain; wherein the first-type biomolecular interaction domain and the second-type biomolecular interaction domain bind specifically to each other; and wherein the fourth-type biomolecular interaction domain and the fifth-type biomolecular interaction domain bind specifically to each other. In some embodiments of any of the aspects, the fourth-type biomolecular interaction domain and fifth-type biomolecular interaction domain have a weaker affinity than the second-type biomolecular interaction domain and first-type protein interaction domain. In some embodiments of any of the aspects, the first polypeptide further comprises a sixth-type biomolecular interaction domain and the recognition polypeptide further comprises a seventh-type biomolecular interaction domain which bind specifically to each other.

In some embodiments of any of the aspects, the first polypeptide comprises the entire TCR recognition domain. In some embodiments of any of the aspects, the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain. In some embodiments of any of the aspects, the TCR recognition domain comprises a non-polypeptide component.

In some embodiments of any of the aspects, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain. In some embodiments of any of the aspects, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.

In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; featureless peptide MHC; or a MHC-peptide fusion. In some embodiments of any of the aspects, the peptide is a human or non-human peptide. In some embodiments of any of the aspects, the peptide is a Minor Histocompatibility Antigen (MiHA). In some embodiments of any of the aspects, the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide complex is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide fusion is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide complex is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC-peptide fusion is a dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In some embodiments of any of the aspects, the MHC is a MHC class I or a MHC class II.

In some embodiments of any of the aspects, the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion. In some embodiments of any of the aspects, the CD1 is CD1d.

In some embodiments of any of the aspects, the biomolecular interaction domains are found on an extracellular portion of the respective polypeptides. In some embodiments of any of the aspects,

-   -   a. wherein the biomolecular interaction domain(s) is a leucine         zipper, or any binding pair of biomolecular interaction domains         are collectively a pair of leucine zippers;     -   b. wherein the biomolecular interaction domain(s) is a BZip (RR)         and/or a AZip (EE), or any binding pair of biomolecular         interaction domains are collectively a BZip (RR) and a AZip         (EE);     -   c. wherein the biomolecular interaction domain(s) is a         PSD95-D1g1-zo-1 (PDZ) domain;     -   d. wherein the biomolecular interaction domain(s) is a         streptavidin and/or a streptavidin binding biomolecular (SBP) or         any binding pair of biomolecular interaction domains are         collectively a streptavidin and a streptavidin binding         biomolecular (SBP);     -   e. wherein the biomolecular interaction domain(s) is a         FKBP-binding domain of mTOR (FRB) and/or a FK506 binding         biomolecular (FKBP) or any binding pair of biomolecular         interaction domains are collectively a FKBP-binding domain of         mTOR (FRB) and a FK506 binding biomolecular (FKBP);     -   f. wherein the biomolecular interaction domain(s) is a         cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506         binding biomolecular (FKBP) or any binding pair of biomolecular         interaction domains are collectively a cyclophilin-Fas fusion         biomolecular (CyP-Fas) and a FK506 binding biomolecular (FKBP);     -   g. wherein the biomolecular interaction domain(s) is a         calcineurin A (CNA) and/or a FK506 binding biomolecular (FKBP)         or any binding pair of biomolecular interaction domains are         collectively a calcineurin A (CNA) and a FK506 binding         biomolecular (FKBP);     -   h. wherein the biomolecular interaction domain(s) is a         gibberellin insensitive (GIA) and/or a gibberellin insensitive         dwarf1 (GID1) or any binding pair of biomolecular interaction         domains are collectively a gibberellin insensitive (GIA) and a         gibberellin insensitive dwarf1 (GID1);     -   i. wherein the biomolecular interaction domain(s) is a Snap-tag         and/or a Halo tag, or any binding pair of biomolecular         interaction domains are collectively a Snap-tag and a Halo tag;     -   j. wherein the biomolecular interaction domain(s) is a         T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or         any binding pair of biomolecular interaction domains are         collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2         (CT52);     -   k. wherein the biomolecular interaction domain(s) is a PYL         and/or a ABI, or any binding pair of biomolecular interaction         domains are collectively a PYL and a ABI;     -   l. wherein the biomolecular interaction domain(s) is a         nucleotide tag and/or a zinc finger domain, or any binding pair         of biomolecular interaction domains are collectively a         nucleotide tag and a zinc finger domain;     -   m. wherein the biomolecular interaction domain(s) is a         nucleotide tag, or any binding pair of biomolecular interaction         domains are collectively a pair of nucleotide tags;     -   n. wherein the biomolecular interaction domain(s) is a         Fluorescein isothiocyanate (FITC) and/or a FITC binding         biomolecular or any binding pair of protein interaction domains         are collectively a FITC and a FITC binding protein; and/or     -   o. wherein the protein interaction domain(s) is a         (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or         any binding pair of protein interaction domains are collectively         a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE binding protein.         In some embodiments of any of the aspects, the nucleotide tag is         a DNA tag or dsDNA tag.

In some embodiments of any of the aspects, the intracellular signaling domain comprises or is a signaling domain from one or more proteins selected from the group consisting of: TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.

In one aspect of any of the embodiments, described herein is a cell comprising and/or expressing a composition described herein. In one aspect of any of the embodiments, described herein is a composition comprising a first polypeptide of any of the preceding claims and a cell expressing or comprising the signaling polypeptide of any of the preceding claims.

In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any of the aspects, the TCR recognition domain comprises a synthetic MHC. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogeneic to the cell. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.

In some embodiments of any of the aspects, the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell. In some embodiments of any of the aspects, the cell is engineered to express one of more of the polypeptide(s) of the composition. In some embodiments of any of the aspects, the cell is engineered to express the signaling polypeptide of the composition. In some embodiments of any of the aspects, the cell is further engineered to knockout or knockdown the native MHCI/II. In some embodiments of any of the aspects, the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.

In one aspect of any of the embodiments, described herein is a composition, comprising a TCR recognition domain and an intracellular signaling domain but not comprising a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain). In one aspect of any of the embodiments, described herein is a composition, comprising a TCR recognition domain and a biomolecular interaction domain (e.g., a first-type biomolecular interaction domain) and but not comprising an intracellular signaling domain.

In one aspect of any of the embodiments, described herein is a method of treating or preventing an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD in a subject in need thereof, the method comprising administering to the subject a composition and/or cell of any of the preceding claims. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC allogeneic to the subject.

In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC autologous to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC xenogeneic to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic to the subject. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells. In some embodiments of any of the aspects, the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is autologous to the transplant cells. In some embodiments of any of the aspects, the transplant is any human or non-human cell, tissue, or organ. In some embodiments of any of the aspects, the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.

In some embodiments of any of the aspects, the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma.

In some embodiments of any of the aspects, the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma. In some embodiments of any of the aspects, the autoimmune disease is thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperproliferative skin diseases, psoriasis such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and psoriasis of the nails, atopy including atopic diseases such as hay fever and Job's syndrome, dermatitis including contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, non-specific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic intraocular inflammatory diseases, urticaria such as chronic allergic urticaria and chronic idiopathic urticaria, including chronic autoimmune urticaria, myositis, polymyositis/dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic scleroderma), sclerosis such as systemic sclerosis, multiple sclerosis (MS) such as spino-optical MS, primary progressive MS (PPMS), and relapsing remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, sclerosis disseminata, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (for example, Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis such as ulcerative colitis, colitis ulcerosa, microscopic colitis, collagenous colitis, colitis polyposa, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), bowel inflammation, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, including adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, an autoimmune hematological disorder, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as in meningitis, herpes gestationis, pemphigoid gestationis, pruritis scroti, autoimmune premature ovarian failure, sudden hearing loss due to an autoimmune condition, IgE-mediated diseases such as anaphylaxis and allergic and atopic rhinitis, encephalitis such as Rasmussen's encephalitis and limbic and/or brainstem encephalitis, uveitis, such as anterior uveitis, acute anterior uveitis, granulomatous uveitis, nongranulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome such as chronic or acute glomerulonephritis such as primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membrano- or membranous proliferative GN (MPGN), including Type I and Type II, and rapidly progressive GN, proliferative nephritis, autoimmune polyglandular endocrine failure, balanitis including balanitis circumscripta plasmacellularis, balanoposthitis, erythema annulare centrifugum, erythema dyschromicum perstans, eythema multiform, granuloma annulare, lichen nitidus, lichen sclerosus et atrophicus, lichen simplex chronicus, lichen spinulosus, lichen planus, lamellar ichthyosis, epidermolytic hyperkeratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reaction, eczema including allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and vesicular palmoplantar eczema, asthma such as asthma bronchiale, bronchial asthma, and auto-immune asthma, conditions involving infiltration of T cells and chronic inflammatory responses, immune reactions against foreign antigens such as fetal A-B-O blood groups during pregnancy, chronic pulmonary inflammatory disease, autoimmune myocarditis, leukocyte adhesion deficiency, lupus, including lupus nephritis, lupus cerebritis, pediatric lupus, non-renal lupus, extra-renal lupus, discoid lupus and discoid lupus erythematosus, alopecia lupus, systemic lupus erythematosus (SLE) such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and lupus erythematosus disseminatus, juvenile onset (Type I) diabetes mellitus, including pediatric insulin-dependent diabetes mellitus (IDDM), adult onset diabetes mellitus (Type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic large-artery disorder, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, sarcoidosis, granulomatosis including lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitides, including vasculitis, large-vessel vasculitis (including polymyalgia rheumatica and giant-cell (Takayasu's) arteritis), medium-vessel vasculitis (including Kawasaki's disease and polyarteritis nodosa/periarteritis nodosa), microscopic polyarteritis, immunovasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, necrotizing vasculitis such as systemic necrotizing vasculitis, and ANCA-associated vasculitis, such as Churg-Strauss vasculitis or syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs positive anemia, Diamond Blackfan anemia, hemolytic anemia or immune hemolytic anemia including autoimmune hemolytic anemia (AIHA), pernicious anemia (anemia perniciosa), Addison's disease, pure red cell anemia or aplasia (PRCA), Factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases involving leukocyte diapedesis, CNS inflammatory disorders, multiple organ injury syndrome such as those secondary to septicemia, trauma or hemorrhage, antigen-antibody complex-mediated diseases, anti-glomerular basement membrane disease, anti-phospholipid antibody syndrome, allergic neuritis, Behcet's disease/syndrome, Castleman's syndrome, Goodpasture's syndrome, Reynaud's syndrome, Sjogren's syndrome, Stevens-Johnson syndrome, pemphigoid such as pemphigoid bullous and skin pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, pemphigus mucus-membrane pemphigoid, and pemphigus erythematosus), autoimmune polyendocrinopathies, Reiter's disease or syndrome, an immune complex disorder such as immune complex nephritis, antibody-mediated nephritis, polyneuropathies, chronic neuropathy such as IgM polyneuropathies or IgM-mediated neuropathy, and autoimmune or immune-mediated thrombocytopenia such as idiopathic thrombocytopenic purpura (ITP) including chronic or acute ITP, scleritis such as idiopathic cerato-scleritis, episcleritis, autoimmune disease of the testis and ovary including autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis such as autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis), or subacute thyroiditis, idiopathic hypothyroidism, Grave's disease, polyglandular syndromes such as autoimmune polyglandular syndromes (or polyglandular endocrinopathy syndromes), paraneoplastic syndromes, including neurologic paraneoplastic syndromes such as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis such as allergic encephalomyelitis or encephalomyelitis allergica and experimental allergic encephalomyelitis (EAE), myasthenia gravis such as thymoma-associated myasthenia gravis, cerebellar degeneration, neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome (OMS), and sensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome, autoimmune hepatitis, lupoid hepatitis, giant-cell hepatitis, autoimmune chronic active hepatitis, lymphoid interstitial pneumonitis (LIP), bronchiolitis obliterans (non-transplant) vs NSIP, Guillain-Barre syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis such as primary biliary cirrhosis and pneumonocirrhosis, autoimmune enteropathy syndrome, Celiac or Coeliac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, amylotrophic lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear disease such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsed or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome/nonsyphilitic interstitial keratitis, Bell's palsy, Sweet's disease/syndrome, rosacea autoimmune, zoster-associated pain, amyloidosis, a non-cancerous lymphocytosis, a primary lymphocytosis, which includes monoclonal B cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndrome, channelopathies including channelopathies of the CNS, autism, inflammatory myopathy, focal or segmental or focal segmental glomerulosclerosis (FSGS), endocrine opthalmopathy, uveoretinitis, chorioretinitis, autoimmune hepatological disorder, fibromyalgia, multiple endocrine failure, Schmidt's syndrome, adrenalitis, gastric atrophy, presenile dementia, demyelinating diseases such as autoimmune demyelinating diseases and chronic inflammatory demyelinating polyneuropathy, Dressler's syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility, e.g., due to anti-spermatozoan antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortion, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, bird-fancier's lung, allergic granulomatous angiitis, benign lymphocytic angiitis, Alport's syndrome, alveolitis such as allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reaction, Sampter's syndrome, Caplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial lung fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilic faciitis, Shulman's syndrome, Felty's syndrome, cyclitis such as chronic cyclitis, heterochronic cyclitis, iridocyclitis (acute or chronic), or Fuch's cyclitis, Henoch-Schonlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndromes, Evan's syndrome, autoimmune gonadal failure, Sydenham's chorea, post-streptococcal nephritis, thromboangitis ubiterans, thyrotoxicosis, tabes dorsalis, chorioiditis, giant-cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplant organ reperfusion, retinal autoimmunity, aphthae, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Boeck's disease, enteritis allergica, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, febris rheumatica, Hamman-Rich's disease, sensoneural hearing loss, ileitis regionalis, leucopenia, transverse myelitis, primary idiopathic myxedema, ophthalmia symphatica, polyradiculitis acuta, pyoderma gangrenosum, acquired spenic atrophy, vitiligo, toxic-shock syndrome, conditions involving infiltration of T cells, leukocyte-adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, diseases involving leukocyte diapedesis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathies, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine failure, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid, frontal, maxillary, or sphenoid sinusitis, an eosinophil-related disorder such as eosinophilia, pulmonary infiltration eosinophilia, eosinophilia-myalgia syndrome, Loffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, granulomas containing eosinophils, seronegative spondyloarthritides, polyendocrine autoimmune disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia telangiectasia syndrome, angiectasis, autoimmune disorders associated with collagen disease, rheumatism, allergic hypersensitivity disorders, glomerulonephritides, reperfusion injury, ischemic re-perfusion disorder, lymphomatous tracheobronchitis, inflammatory dermatoses, dermatoses with acute inflammatory components, and autoimmune uveoretinitis (AUR).

In some embodiments of any of the aspects, the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy. In some embodiments of any of the aspects, the biologic, cell-therapy, or gene therapy is an adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.

In some embodiments of any of the aspects, the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. In some embodiments of any of the aspects, the disease is vitiligo and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO: 2021. In some embodiments of any of the aspects, the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 80% or at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO: 2039. In some embodiments of any of the aspects, the disease is type 1 diabetes and the TCR recognition domain comprises one or more of SEQ ID NOs: 8-17; comprises HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or comprises HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129. In some embodiments of any of the aspects, the disease is vitiligo and the TCR recognition domain comprises SEQ ID NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO: 2018; or comprises HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or comprises HLA-A*0101 and SEQ ID NO: 2021. In some embodiments of any of the aspects, the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or comprises HLA-B*0702 and SEQ ID NO: 2038; or comprises HLA-B*0801 and SEQ ID NO: 2039.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 demonstrates that pMHC tetramer+FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion or manner. I-H2Kb: MHC class I tetramer. I-Ab-: Control tetramer. Figure discloses SEQ ID NOS 2752-2753, 2750 and 2754, respectively, in order of appearance.

FIG. 2 depicts expression of activation marker CD69 on OTi cells at different time points (24, 48, 72 hrs.). Binding of Jurkat+ pMHC to OTi cells does not change Jurkat live count (right). Figure discloses SEQ ID NOS 2750, 2750 and 2754, respectively, in order of appearance.

FIG. 3 depicts the expression of CD69 on Jurkat CAR, e.g., CAL T cells in different time points (24, 48, 72 hrs.). Figure discloses SEQ ID NOS 2750 and 2755, respectively, in order of appearance.

FIG. 4 depicts the cytotoxicity of Primary human CD8 CAR, e.g., CAL T cells with different concentrations of tetramers (left). Minimal activation of target cells (CD69 on CAR, e.g., CAL T cells and target cells) after coculturing with OTi cells (FIG. 2 left). I-Ab: Ctrl/H2Kb: MHC-I. Figure discloses SEQ ID NOS 2750, 2754 and 2750, respectively, in order of appearance.

FIG. 5 demonstrates that the cytotoxicity of human CD8 CAR, e.g., CAL T is highly specific and was not seen with ctrl tetramer (left). No cytotoxicity was seen on killer CAR, e.g., CAL T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes. Figure discloses SEQ ID NOS 2754, 2750, 2754, 2750, 2754 and 2750, respectively, in order of appearance.

FIG. 6A depicts a schematic of SUPRA CAR design applied to provide Universal CAL. This design separates the cell targeting molecule module from the killer cell. FIG. 6B depicts that SUPRA CARs can be designed with CD3 domain uncoupled from the costimulatory domains to provide Universal CAL as described herein.

FIGS. 7A-7D depict key features of the SUPRA CAR systems that can be applied to Universal CAL. FIG. 7A demonstrates that zipCAR activation is tunable through modulation of zipFv concentration, zipper affinity, scFv affinity, and zipCAR expression level in human primary CD4 T cells, as demonstrated by IFN-g production. FIG. 7B demonstrates that SUPRA CAR system, as applied to Universal CAL, can perform combination antigen detection to form AND gate logic in CD4 T cells. FIG. 7C demonstrates that xenograft animal tumor model shows tumor eradication (as demonstrated by luciferase photon flux given by the tumor cells) by the SUPRA CAR T cells. FIG. 7D demonstrates that SUPRA CARs as applied to Universal CAL can be used to control different cell types, such as CD4 and CD8 T cells, against two different antigens. CD69 expression (a T cell activation marker) is quantified with flow cytometry for CD4 and CD8 T cells. (From Cho, Collins, and Wong, Cell. 2018)

FIG. 8 depicts the timeline of double Hu-PBMC-HSCT-skin graft mouse model generation.

FIG. 9 depicts a summary of double hu-PBMC-HSCT-skin graft mouse model generation.

FIG. 10 depicts key features of pMHC multimer+CAR, e.g., CAL T cell system.

FIG. 11 depicts a table of experimental design. Figure discloses SEQ ID NOS 2750, 2756 and 2754, respectively, in order of appearance.

FIG. 12 demonstrates verification of FITC-conjugated tetramer mediated activation. Figure discloses SEQ ID NOS 2750 and 2751, respectively, in order of appearance.

FIG. 13 depicts a time course of FITC-conjugated tetramer mediated activation. Figure discloses SEQ ID NOS 2750, 2751, 2750, 2751, 2750, 2751, 2750 and 2751, respectively, in order of appearance.

FIG. 14 depicts a graph of Jurkat cell counts. Figure discloses SEQ ID NOS 2750 and 2754, respectively, in order of appearance.

FIG. 15 depicts tetramer staining. Figure discloses SEQ ID NOS 2750, 2754, 2751, 2756, 2754, 2750 and 2754, from left to right and top to bottom.

FIG. 16 depicts heatmaps of indicated staining levels. Figure discloses SEQ ID NOS 2751, 2757, 2754, 2750, 2754, 2751, 2754, 2757, 2750, 2754, 2751, 2754, 2757, 2750 and 2754, respectively, in order of appearance.

FIG. 17 depicts a table of experimental design.

FIGS. 18-20 depict graphs of cytotoxicity levels. FIG. 18 discloses SEQ ID NOS 2754, 2750, 2754 and 2750 from left to right. FIG. 19 discloses SEQ ID NOS 2754 and 2750, respectively, in order of appearance. FIG. 20 discloses SEQ ID NOS 2750, 2754, 2750 and 2754 from left to right.

FIG. 21 depicts the levels of CD69 on OTi CD8 T cells. Figure discloses SEQ ID NOS 2750, 2754, 2750 and 2754 from left to right.

FIG. 22 depicts schematics of two embodiments of the technology described herein.

FIG. 23 depicts a schematic of the FU-CAL embodiments of the technology described herein.

FIG. 24 depicts a schematic of the CAL-BITE embodiments of the technology described herein. The left panel depicts blinatumomab, which is described in more detail in Weiner et al. The Molecular Basis of Cancer 2015 683-694.e3.

FIG. 25 depicts a schematic of the CAL technology disarming autoreactive T cells.

FIGS. 26A-26B depict schematics of T cells design. MHC can be mouse or human. The MiHA can be ovalbumin (against OTi or OTii) or disparate antigens between donor and recipient.

FIG. 27 depicts a graph demonstrating that pMHC tetramer+FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion, while OTii specific tetramer does not.

FIG. 28A depicts expression of CD69 on OTi cells at different time points (24, 48, 72 hrs.).

FIG. 28B is a graph demonstrating that binding of Jurkat+ pMHC to OTi cells does not change Jurkat live count.

FIG. 29 depicts a graph demonstrating that cytotoxicity of pMHC-CAR against 1E6 T cell clone exhibits a dose dependent association.

FIG. 30 depicts a graph demonstrating that CAR Jurkat T cells activation (as measured by CD69 surface marker) after exposure to target cells (OTi or OTii TCR expressing T cells) and the corresponding tetramer (OTi or OTii specific) (n=4).

FIG. 31 depicts graphs demonstrating that cytotoxicity of human CD8 CAR T is highly specific and was not seen with ctrl tetramer. No cytotoxicity was seen on killer CAR T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes (n=4).

FIG. 32 depicts a graph demonstrating cytotoxicity of Primary CD8 CAR T cells against OTi TCR T cells with different concentration of tetramer.

DETAILED DESCRIPTION

Aspects of the invention described herein relate to chimeric antigen ligands. As used herein, “chimeric antigen ligand” or “CAL” refers to an artificially constructed molecule comprising a TCR recognition domain (e.g. an polypeptide comprising at least one MHC sequence as described herein) and at least one biomolecular interaction domain. The TCR recognition domain is selected to bind to specific populations of T cells that it is desirable to target and/or destroy, e.g., for therapeutic purposes in T-cell mediated diseases. In some embodiments, the population of targeted T cells is a population of polyclonal pathogenic T cells. The biomolecular interaction domain is selected to bind to a second cell, e.g., a NK cell, thereby colocalizing the targeted T cell and the second cell and promoting or increasing the inhibition and/or destruction of the targeted T cell. In various embodiments, the CAL is selected to have high affinity or avidity for the TCR, e.g., the TCR variable domain.

The CALs can be used herein with endogenous cells, e.g., in some embodiments, no engineered cells are administered to the subject. In other embodiments, the CALs can be used with engineered cells, e.g., engineered NK cells. In such cases, the engineered cells can comprise one or more CARs, e.g., a CAR comprising an extracellular domain with a biomolecular interaction domain that specifically binds with the biomolecular interaction domain of the CAL.

Accordingly, described herein are chimeric antigen receptors (CARs) in which the recognition and signaling portions of the CAR are separate polypeptides. The two separate polypeptides that make up a complete CAR are able to interact and form the complete CAR by way of protein interaction domains. This permits flexible, modular CAR-T therapy which is capable of complex logic computation, providing a more precise and effective approach to immunotherapy.

In one aspect of any of the embodiments is a CAL and/or chimeric antigen receptor (CAR) having multiple components, and/or a cell or composition comprising a multi-component CAL and/or CAR. Multi-component CALs/CARs are also referred to herein variously as SMART CAL/CAR or SUPRA.

As used herein, traditional “chimeric antigen receptor” or “CAR” refers to an artificially constructed hybrid polypeptide comprising an antigen-binding domain (e.g. an antigen-binding portion of an antibody (e.g. a scFV)) linked to a cell signaling and/or cell activation domain. In some embodiments the cell-signaling domain can be a T-cell signaling domain. In some embodiments, the cell activation domain can be a T-cell activation domain. CARs have the ability to redirect the specificity and reactivity of T cells and other immune cells (e.g., NK cells) toward a selected target in a non-MHC-restricted manner, exploiting the antigen-binding properties of monoclonal antibodies.

The non-MHC-restricted antigen recognition gives T-cells expressing CARs the ability to recognize an antigen independent of antigen processing, thus bypassing a major mechanism of tumor escape. Moreover, when expressed in T-cells, CARs advantageously do not dimerize with endogenous T-cell receptor (TCR) alpha and beta chains. Most commonly, the CAR's extracellular binding domain is composed of a single chain variable fragment (scFv) derived from fusing the variable heavy and light regions of a murine or humanized monoclonal antibody. Alternatively, scFvs may be used that are derived from Fabs (instead of from an antibody, e.g., obtained from Fab libraries), in various embodiments, this scFv is fused to a transmembrane domain and then to an intracellular signaling domain. “First-generation” CARs include those that solely provide CD3zeta signals upon antigen binding, “Second-generation” CARs include those that provide both costimulation (e.g. CD28 or CD 137) and activation (CD3Q). “Third-generation” CARs include those that provide multiple costimulation (e.g. CD28 and CD 137) and activation (CO3Q). In various embodiments, the CAR is selected to have high affinity or avidity for the antigen. Further discussion of CARs can be found, e.g., in Maus et al. Blood 2014 123:2624-35; Reardon et al. Neuro-Oncology 2014 16:1441-1458; Hoyos et al. Haematologica 2012 97:1622; Byrd et al. J Clin Oncol 2014 32:3039-47; Maher et al. Cancer Res 2009 69:4559-4562; and Tamada et al. Clin Cancer Res 2012 18:6436-6445; each of which is incorporated by reference herein in its entirety.

As used herein, “multi-component CAL” refers to a CAL comprising at least two separate polypeptides, neither of which polypeptides is capable of both ligand recognition and signaling activation on its own. As used herein, “multi-component CAR” refers to a CAR comprising at least two separate polypeptides, neither of which polypeptides is capable of both ligand recognition and signaling activation on its own. In some embodiments, the at least two separate polypeptides each comprise a protein interaction domain that permits interaction, e.g., binding of the separate polypeptides. In some embodiments, one of the at least two separate polypeptides is a transmembrane polypeptide having an intracellular T cell receptor (TCR) signaling domain and a second of the at least two separate polypeptides is an extracellular polypeptide having a ligand-binding domain. In some embodiments, a multi-component CAL and/or CAR can comprise two, three, four, five, six, seven, eight, nine, ten or more separate polypeptides.

Various aspects provided herein provide a composition comprising multiple components of a multi-component CAL and/or CAR.

In one aspect of the embodiments is a composition, e.g., a single molecule, comprising a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain. In one aspect of the embodiments is a composition, e.g., a single molecule comprising a TCR recognition domain; and a first-type biomolecular (e.g., protein) interaction domain. Further provided herein is a multi-component CAL and/or CAR comprising a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain. In some embodiments, the composition, e.g, single molecule, comprising a TCR recognition domain; and a first-type biomolecular (e.g., protein) interaction domain does not comprise an antibody, antibody domain, or antibody reagent.

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other. Further provided herein is a multi-component CAL and/or CAR comprising a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other. Further provided herein is a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other.

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain. Further provided herein is a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.

In various embodiments, the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain Affinity can be measured by one skilled in the art using standard methods, for example, by measuring its equilibrium dissociation constant (K_(d)).

Another aspect of the embodiments is a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other. Further provided herein is a multi-component CAL and/or CAR comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

In various embodiments, the fourth-type protein interaction domain and fifth-type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain. Affinity can be measured as described above.

In various embodiments, the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.

In some embodiments, the first polypeptide comprises the entire TCR recognition domain. In some embodiments, the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.

In some embodiments, a composition described herein can comprise multiple copies or instances of a TCR recognition domain(s), e.g. the TCR recognition domain can be a mulitmer, or oligomer. In some embodiments, a composition described herein can comprise multiple copies or instances of a first polypeptide as described herein.

In various embodiments, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain. In one embodiment, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.

As used herein, “TCR recognition domain” refers to a domain or portion of a polypeptide that can target or bind specifically to a TCR, e.g., a TCR expressed on the surface of T cell. In some embodiments, the TCR recognition domain can be a TCR variable region (TCR-VR) recognition domain, i.e. is can target or bind specifically to the variable region of a TCR, e.g., a TCR expressed on the surface of a T cell. The TCR recognition domain sequence can be autologous, allogeneic, or xenogeneic to a given subject. In some embodiments, the TCR recognition domain sequence is a wild-type protein or sequence. In some embodiments, the TCR recognition domain sequence is a naturally-occurring variant, e.g., an allele of a wild-type protein or sequence. In some embodiments, the TCR recognition domain sequence is modified relative to a wild-type protein, e.g. chemically modified. In some embodiments, the TCR recognition domain sequence is a derivative and/or variant of a wild-type sequence. In some embodiments, the TCR recognition domain sequence can be a human, or non-human sequence.

A TCR recognition domain can comprise a MHC polypeptide, a MHC polypeptide sequence, and/or comprise a portion of a MHC sequence. The MHC and/or MHC sequence can be autologous, allogeneic, or xenogeneic to a given subject. In some embodiments, the MHC and/or MHC sequence is a wild-type protein or sequence. In some embodiments, the MHC and/or MHC sequence is a naturally-occurring variant, e.g., an allele of MHC. In some embodiments, the MHC and/or MHC sequence is modified relative to a wild-type protein, e.g. chemically modified. In some embodiments, the MHC and/or MHC sequence is a derivative and/or variant of a wild-type MHC sequence. In some embodiments, the MHC and/or MHC sequence can be or comprise a human, or non-human sequence.

In one embodiment, the TCR recognition domain comprises a MHC (Major Histocompatibility Complex), a MHC-peptide complex, or a MHC-peptide fusion. In some embodiments, the TCR recognition domain can comprise a featureless peptide MHC, or a MHC without peptides, or any other molecule that can target or bind specifically to the variable region of the TCR.

The MHC, which is also referred to as the human leukocyte antigen (HLA), is comprised of a set of genes that code for cell surface proteins essential for the acquired. e.g., adaptive immune system to recognize foreign molecules in vertebrates, which in turn determines histocompatibility. The MHC gene family is divided into three subgroups: MHC class I, MHC class II, and MHC class III. Class I MHC molecules have the β2 microglobulin subunit which can only be recognised by CD8 co-receptors. Class II MHC molecules have β1 and β2 subunits and can be recognized by CD4 co-receptors. In this way MHC molecules chaperone, which type of lymphocytes bind to the given antigen with high affinity, since different lymphocytes express different T-Cell Receptor (TCR) co-receptors. Components of the MHC are known in the art and can be readily identified by a skilled person. The MHC is further described in, e.g., Janeway C A Jr, Travers P, Walport M, et al, Immunobiology: The Immune System in Health and Disease, 5th edn (New York: Garland Science, 2001); Vigneron N, Stroobant V, Chapiro J, Ooms A, Degiovanni G, Morel S, et al. (April 2004). “An antigenic peptide produced by peptide splicing in the proteasome”. Science. 304 (5670): 587-90; and K. Murphy, “Antigen recognition by T cells,” in Janeway's Immunobiology, 8th, Ed., Garland Science, 2012, pp. 138-153; which are incorporated herein by reference in their entireties. A complete MHC class I complex comprises one MHC class I heavy chain, one peptide ligand sequence, and a beta 2 microglobulin. In some embodiments, a TCR recognition domain comprises one MHC class I heavy chain, one peptide ligand sequence, and a beta 2 microglobulin. A complete MHC class II complex comprises an MHC class II alpha chain, MHC class II beta chain, and one peptide ligand sequence. In some embodiments, a TCR recognition domain comprises an MHC class II alpha chain, MHC class II beta chain, and one peptide ligand sequence. These three components can either be assembled together as separate sequences (e.g., by intramolecular binding of multiple peptide molecules) or can be expressed as fusion proteins with intervening linker sequences (e.g., see Schmittnaegel et al., 2016; which is incorporated by reference herein in its entirety).

The MHC (Major Histocompatibility Complex), MHC-peptide complex, MHC-peptide fusion, featureless peptide MHC, can be selected on the basis of the disease or condition to be treated/prevented. Specific MHCs, peptides, and/or antigens that are associated with the diseases described herein are known in the art and an appropriate MHC, peptide, and/or antigen can be selected by one of ordinary skill in the art. For example, databases of suitable MHC, peptide, and/or antigen sequences are available on the world wide web at iedb.org; immunespace.org; immgen.org; import/org; peptideatlast.org/repository/; uniprot.org; ncbi.nlm.nih.gov/protein/; immunedata.org/index.php; immuneprofiling.org/hipc/; allergenonline.org/databasebrowe.shtml; and itntrialshare.org. Further examples are also provided in Smatti et al. 2019 Viruses 11:762; Beretta-Piccoli et al. 2019 J Autoimmu 94:1-6; and Cusick et al. 2012 Clinical Reviews in Allergy and Immunology; each of which is incorporated by reference herein in its entirety. The specific examples of TCR recognition domains provides herein are exemplary and non-limiting. One of skill in the art can identify relevant auto-antigenic pMHCs, allogeneic peptide MHCs, and autogenic peptide MHCs in addition to those described herein, e.g., from the art and/or from donor cells. Such identification is within the skill of the ordinary practitioner.

As an illustrative and non-limiting example, for compositions and methods relating to type 1 diabetes, the TCR recognition domain can comprise one or more of SEQ ID NOs: 8-17. In some embodiments, the TCR recognition domain can comprise SEQ ID NO: 8, 9, and one of 10-17. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to SEQ ID NO: 8, 9, and one of 10-17. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to SEQ ID NO: 8, 9, and one of 10-17, and which retain the wild-type activity of SEQ ID NOs: 8, 9, and one of 10-17.

HLA-A*0201 (MHC class Iheavy chain allele, wildtype, human) SEQ ID NO: 8 MAVMAPRTLVLLLSGALALTQTWAGSHSMRYFFTSVSRPGRGEPRFIAVG YVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGETRKVKAHSQTHRV DLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDYIAL KEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGK ETLQRTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQ DTELVETRPAGDGTFQKWAAVVVPSGQEQRYTCHVQHEGLPKPLTLRWEP SSQPTIPIVGIIAGLVLFGAVITGAVVAAVMWRRKSSDRKGGSYSQAASS DSAQGSDVSLTACKV Beta-2 microglobulin (wildtype, human) SEQ ID NO: 9 MSRSVALAVLALLSLSGLEGIQRTPKIQVYSRHPAENGKSNFLNCYVSGF HQSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEYAC RVNHVTLSQPKIVKWDRDM Preproinsulin₁₅₋₂₄ (wildtype, human) SEQ ID NO: 10 ALWGPDPAAA Preproinsulin₁₅₋₂₄ altered peptide ligand #1 (synthetic, Cole et al., 2016) SEQ ID NO: 11 AQWGPDPAAA Preproinsulin₁₅₋₂₄ altered peptide ligand #2 (synthetic, Cole et al., 2016) SEQ ID NO: 12 RQWGPDPAAV Preproinsulin₁₅₋₂₄ altered peptide ligand #3 (wildtype, Clostridium asparagiforme, Cole et al., 2016) SEQ ID NO: 13 RQFGPDWIVA Preproinsulin₁₅₋₂₄ altered peptide ligand #4 (synthetic, Cole et al., 2016) SEQ ID NO: 14 YQFGPDFPIA Preproinsulin₁₅₋₂₄ altered peptide ligand #5 (synthetic, Cole et al., 2016) SEQ ID NO: 15 RQFGPDFPTI Preproinsulin₁₅₋₂₄ altered peptide ligand #6 (synthetic, Cole et al., 2016) SEQ ID NO: 16 YLGGPDFPTI Preproinsulin₁₅₋₂₄ altered peptide ligand #7 (wildtype, Bacteroides fragilis, Cole et al., 2016) SEQ ID NO: 17 MVWGPDLYV

As a further illustrative example, for compositions and methods relating vitiligo, the TCR recognition domain can comprise one or more of SEQ ID NOs: 18-22. In some embodiments, the TCR recognition domain can comprise SEQ ID NO: 18, 19, and one of 20-22. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to SEQ ID NO: 18, 19, and one of 20-22. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to SEQ ID NO: 18, 19, and one of 20-22, and which retain the wild-type activity of SEQ ID NOs: 18, 19, and one of 20-22.

HLA-A*0201 (MHC class I heavy chain allele, wildtype, human, same as previous HLA-A2) SEQ ID NO: 18 MAVMAPRTLVLLLSGALALTQTWAGSHSMRYFFTSVSRPGRGEPRFIAVG YVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGETRKVKAHSQTHRV DLGTLRGYYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDYIAL KEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGK ETLQRTDAPKTHMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQ DTELVETRPAGDGTFQKWAAVVVPSGQEQRYTCHVQHEGLPKPLTLRWEP SSQPTIPIVGIIAGLVLFGAVITGAVVAAVMWRRKSSDRKGGSYSQAASS DSAQGSDVSLTACKV Beta-2 microglobulin (wildtype, human, same as previous Beta-2 microglobulin) SEQ ID NO: 19 MSRSVALAVLALLSLSGLEGIQRTPKIQVYSRHPAENGKSNFLNCYVSGF HQSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEYAC RVNHVTLSQPKIVKWDRDM MART-1₂₆₋₃₅ (wildtype, human) SEQ ID NO: 20 ELAGIGILTV Tyrosinase₃₆₈₋₃₇₆ (wildtype, human) SEQ ID NO: 21 YMDGTMSQV gp100₂₀₉₋₂₁₇ (wildtype, human) SEQ ID NO: 22 ITDQVPFSV

As further illustrative and non-limiting examples, the following pairs of MHC and antigens provided in Table 5 are known in the art. In some embodiments, the TCR recognition domain can comprise one or more of the following indicated MHC/peptide pairs, e.g., the TCR recognition domain can comprise one of the indicated MHC alleles and the indicated corresponding peptide. In some embodiments, the TCR recognition domain can comprise sequences with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of the following indicated MHC/peptide pairs, e.g., one of the indicated MHC alleles and the indicated corresponding peptide. In some embodiments, the TCR recognition domain can comprise sequences with at least 95% sequence identity to one of the following indicated MHC/peptide pairs, e.g., one of the indicated MHC alleles and the indicated corresponding peptide, wherein those sequences retain the wild-type activity of the MHC allele and the corresponding peptide.

TABLE 5 Each line of the following Table provides a MHC allele (whose sequence is available in publically-accessible databases, e.g., (NCBI), and an antigen sequence. The antigen source and/or the relevant disease are also indicated on some lines. In some examples provided in Table 5, the peptides are antigen mimics and their origin is indicated. HLA-A*0201 VMNILLQYVV GAD65 Diabetes (SEQ ID NO: 2013) HLA-A*0201 VMNILLQYVV GAD65 Diabetes (SEQ ID NO: 2013) HLA-A*0201 YAYDGKDYIA HLA-A2 Diabetes (SEQ ID NO: 2014) HLA-A*0201 MVWESGCTV IA-2 Diabetes (SEQ ID NO: 2015) HLA-A*0201 VIVMLTPLV IA-2 Diabetes (SEQ ID NO: 2016) HLA-A*0201 YTCPLCRAPV SAA Autoimmune (SEQ ID NO: 2017) HLA-A*0201 YMDGTMSQV Tyrosinase Vitiligo (SEQ ID NO: 2018) HLA-A*0301 YMVPFIPLYR Tyrosinase Vitiligo (SEQ ID NO: 2019) HLA-A*2402 AFLPWHRLF Tyrosinase Vitiligo (SEQ ID NO: 2020) HLA-A*0101 SSDYVIPIGTY Tyrosinase Vitiligo (SEQ ID NO: 2021) HLA-A*02: VLHDDLLEA HA-1 137-145 Minor Histocompatibility Antigen 01 (SEQ ID NO: 2022) HLA-A*02: RTLDKVLEV miHAg HA-8 Minor Histocompatibility Antigen 01 (SEQ ID NO: 2023) HLA-A*02: FIDSYICQV miHAg H-Y (human SMCY) 311-319 Minor Histocompatibility 01 (SEQ ID NO: 2024) Antigen DRB1*04: 01 GAGSLQPLALEGSLQKRG A Proinsulin 73-90 (SEQ ID NO: 2025) DRB1*04: 01 IAFTSEHSHFSLK A GAD65 274-286 (SEQ ID NO: 2026) DRB1*04: 01 DENPVVHFFKNIVTPRTPP Myelin basic protein 83-101 (SEQ ID NO: 2027) DRB1*04: 01 GIVEQCCTSICSLYQ A Proinsulin 90-104 (SEQ ID NO: 2028) DRB1*04: 01 NFIRMVISNPAAT A GAD65 555-567 (SEQ ID NO: 2029) DRB1*04: 01 DVMNILLQYVVKSFDRSTKV GAD65 13-132 (SEQ ID NO: 2030) HLA-A*0201 ALWGPDPAAA Insulin Diabetes (SEQ ID NO: 2031) HLA-A*0201 HLVEALYLV Insulin Diabetes (SEQ ID NO: 2032) HLA-A*0201 KLQVFLIVL IAPP Diabetes (SEQ ID NO: 2033) HLA-A*0101 IVDCLTEMY USP9Y Graft vs Host (SEQ ID NO: 2034) HLA-A*0201 VLHDDLLEA HA-1 Graft vs Host (SEQ ID NO: 2035) HLA-A*0201 RTLDKVLEV HA-8 Graft vs Host (SEQ ID NO: 2036) HLA-A*0201 FIDSYICQV H-Y Graft vs Host (SEQ ID NO: 2037) HLA-B*0702 SPSVDKARAEL SMCY Graft vs Host (SEQ ID NO: 2038) HLA-B*0801 LPHNHTDL TPR-protein Graft vs Host (SEQ ID NO: 2039) Antigen Mimicry HLA-A*0201 FLDKGTYTL BALF4 EBV (SEQ ID NO: 2040) HLA-A*0201 GLCTLVAML BMLF1 EBV (SEQ ID NO: 2041) HLA-A*0201 TLDYKPLSV BMRF1 EBV (SEQ ID NO: 2042) HLA-A*0201 YVLDHLIVV BRLF1 EBV (SEQ ID NO: 2043) HLA-A*0201 LLDFVRFMGV EBNA 3B EBV (SEQ ID NO: 2044) HLA-A*0201 YLLEMLWRL LMP-1 EBV (SEQ ID NO: 2045) HLA-A*0201 YLQQNWWTL LMP-1 EBV (SEQ ID NO: 2046) HLA-A*0201 CLGGLLTMV LMP-2A EBV (SEQ ID NO: 2047) HLA-A*0201 FLYALALLL LMP-2A EBV (SEQ ID NO: 2048) HLA-A*0301 RLRAEAQVK EMNA 3A EBV (SEQ ID NO: 2049) HLA-A*1101 AVFDRKSDAK EBNA 3B EBV (SEQ ID NO: 2050) HLA-A*1101 IVTDFSVIK EBNA 3B EBV (SEQ ID NO: 2051) HLA-A*2402 DYCNVLNKEF BRLF1 EBV (SEQ ID NO: 2052) HLA-A*2402 TYGPVFMCL LMP-2 EBV (SEQ ID NO: 2053) HLA-A*2902 IACPIVMRY BRLF1 EBV (SEQ ID NO: 2054) HLA-A*6801 IVTDFSVIK EBNA 3B EBV (SEQ ID NO: 2051) HLA-B*0702 RPQGGSRPEFVKL BMRF1 EBV (SEQ ID NO: 2055) HLA-B*0702 RPPIFIRRL EBNA 3A EBV (SEQ ID NO: 2056) HLA-B*0702 QPRAPIRPI EBNA 6 EBV (SEQ ID NO: 2057) HLA-B*0801 RAKFKQLL BZLF1 EBV (SEQ ID NO: 2058) HLA-B*0801 FLRGRAYGL EBNA 3A EBV (SEQ ID NO: 2059) HLA-B*3501 EPLPQGQLTAY BZLF1 EBV (SEQ ID NO: 2060) HLA-B*3501 EPLSQSQITAY BZLF1 EBV (SEQ ID NO: 2061) HLA-B*3501 HPVAEADYFEY BZLF1 1 EBV (SEQ ID NO: 2062) HLA-B*3501 HPVGDADYFEY EBNA 1 EBV (SEQ ID NO: 2063) HLA-B*3501 HPVGEADYFEY EBNA 1 EBV (SEQ ID NO: 2064) HLA-B*3501 HPVGQADYFEY EBNA 1 EBV (SEQ ID NO: 2065) HLA-B*3501 YPLHEQHGM EBNA 3A EBV (SEQ ID NO: 2066) HLA-A*0101 YTEHDTLLY UL44 CMV (SEQ ID NO: 2067) HLA-A*0201 VLEETSVML IE-1 CMV (SEQ ID NO: 2068) HLA-A*0201 NLVPMVATV pp65 CMV (SEQ ID NO: 2069) HLA-A*0301 KLGGALQAK IE-1 CMV (SEQ ID NO: 2070) HLA-A*2301 QYDPVAALF pp65 CMV (SEQ ID NO: 2071) HLA-A*2402 AYAQKIFKI IE-1 CMV (SEQ ID NO: 2072) HLA-A*2402 QYDPVAALF pp65 CMV (SEQ ID NO: 2071) HLA-A*2402 VYALPLKML pp65 CMV (SEQ ID NO: 2073) HLA-B*0702 RPHERNGFTVL pp65 CMV (SEQ ID NO: 2074) HLA-B*0702 TPRVTGGGAM pp65 CMV (SEQ ID NO: 2075) HLA-B*0801 ELKRKMIYM IE-1 CMV (SEQ ID NO: 2076) HLA-B*0801 ELNRKMIYM IE-1 CMV (SEQ ID NO: 2077) HLA-B*0801 ELRRKMMYM IE-1 CMV (SEQ ID NO: 2078) HLA-B*0801 QIKVRVDMV IE-1 CMV (SEQ ID NO: 2079) HLA-B*3501 IPSINVHHY pp65 CMV (SEQ ID NO: 2080) HLA-B*3501 LPLNVGLPIIGVM UL138 CMV (SEQ ID NO: 2081) A*01: 01 SADNNNSEY AAV VP1 492-500 (SEQ ID NO: 2082) A*01: 01 TDLGQNLLY Adenovirus 5 Hexon 886-894 (SEQ ID NO: 2083) A*01: 01 EADPTGHSY MAGE-A1 161-169 (SEQ ID NO: 2084) A*01: 01 EVDPIGHLY MAGE-A3 168-176 (SEQ ID NO: 2085) A*01: 01 KSDICTDEY Tyrosinase 243-251 (244S) (SEQ ID NO: 2086) A*01: 01 KCDICTDEY Tyrosinase 243-251 (SEQ ID NO: 2087) A*01: 01 QSLEIISRY Mcl-1 177-185 (SEQ ID NO: 2088) A*01: 01 YVDFREYEYY FLT3 ITD (SEQ ID NO: 2089) A*01: 01 TLDTLTAFY Mesothelin 429-437 (SEQ ID NO: 2090) A*01: 01 LTDDRLFTCY PLEKHM2 (SEQ ID NO: 2091) A*01: 01 DSDPDSFQDY Tyr A1a 454-463 (SEQ ID NO: 2092) A*01: 01 EADPIGHLY MAGEA3 (SEQ ID NO: 2093) A*01: 01 EVDPASNTY MAGE-A4 169-177 (SEQ ID NO: 2094) A*01: 01 HSTNGVTRIY PSMA (SEQ ID NO: 2095) A*01: 01 ILDTAGREEY N-ras 55-64 (SEQ ID NO: 2096) A*01: 01 LVDVMPWLQY Cytochrome P450 240-249 (SEQ ID NO: 2097) A*01: 01 RSDSGQQARY AIM-2 (SEQ ID NO: 2098) A*01: 01 VTEPGTAQY Minor antigen HA-3T (Lbc oncogene (SEQ ID NO: 2099) 451-459) A*01: 01 VYDFFVWLHY TRP-2 181-190 (SEQ ID NO: 2100) A*01: 01 YSEHPTFTSQY HCMV pp65 363-373 (SEQ ID NO: 2101) A*01: 01 VTEHDTLLY HCMV pp50 245-253 (SEQ ID NO: 2102) A*01: 01 FTSDYYQLY SARS-CoV-2 ORF3a 207-215 (SEQ ID NO: 2103) (confirmed epitope) A*01: 01 TTDPSFLGRY SARS-CoV-2 Replicase polyprotein lab (SEQ ID NO: 2104) 1637-1646 A*01: 01 PTDNYITTY SARS-CoV-2 Replicase polyprotein lab (SEQ ID NO: 2105) 1621-1629 A*01: 01 LLDTASALY HBV core 30-38 (SEQ ID NO: 2106) A*01: 01 ATDALMTGY HCV NS3 1435-1443 (SEQ ID NO: 2107) A*01: 01 ATDALMTGF HCV NS3 1436-1444 (SEQ ID NO: 2108) A*01: 01 CTELKLSDY Influenza A (PR8) NP 44-52 (SEQ ID NO: 2109) A*01: 01 VSDGGPNLY Influenza A PB1 591-599 (SEQ ID NO: 2110) A*01: 01 IVDCLTEMY DRRFY (1521-1529)) (SEQ ID NO: 2111) A*0201 ALCNTDSPL iLR1 (SEQ ID NO: 2112) A*0201 ALKDVEERV MAGE-C2 336-344 (SEQ ID NO: 2113) A*0201 LLAARAIVAI iLR1 59-68 (SEQ ID NO: 2114) A*0201 RLWQELSDI circadian clock protein PASD1 691-700 (SEQ ID NO: 2115) A*0201 LLFGLALIEV MAGE-C2 191-200 (SEQ ID NO: 2116) A*0201 FLDPRPLTV CYP190 (SEQ ID NO: 2117) A*0201 STLCQVEPV MPP11 (SEQ ID NO: 2118) A*0201 VLQMKEEDV iLR1 (SEQ ID NO: 2119) A*0201 AIQDLCLAV NPM1 (SEQ ID NO: 2120) A*0201 QLLIKAVNL MPP11 (SEQ ID NO: 2121) A*0201 AIQDLCVAV NPM1 (SEQ ID NO: 2122) A*0201 ALTPVVVTL cyclin-dependent kinase 4 170-178 (SEQ ID NO: 2123) A*02: 01 KLQVFLIVL T1D Diabetes human prepro islet (SEQ ID NO: 2124) amyloid polypeptide pplAPP 513 A*02: 01 VMNILLQYV GAD65 114-123 (SEQ ID NO: 2125) A*02: 01 SLSRFSWGA Myelin basic protein 110-118 (SEQ ID NO: 2126) A*02: 01 HLVEALYLV Insulin B chain 10-18 (SEQ ID NO: 2127) A*02: 01 LNIDLLWSV T1D Diabetes IGRP 228-236 (SEQ ID NO: 2128) A*02: 01 VLFGLGFAI T1D Diabetes IGRP 265-273 (SEQ ID NO: 2129) A*02: 01 ALWGPDPAAA Proinsulin precursor 15-24 (SEQ ID NO: 2130) A*02: 01 MVWESGCTV IA-2 797-805 (SEQ ID NO: 2131) A*02: 01 YTCPLCRAPV SSA SS-56 55-64 (SEQ ID NO: 2132) A*02: 01 VIVMLTPLV IA-2 805-813 (SEQ ID NO: 2133) A*02: 01 AITEVECFL VP1 44-52 (SEQ ID NO: 2134) A*02: 01 FLHCIVFNV large Tantigen 410-418 (SEQ ID NO: 2135) A*02: 01 LLMWEAVTV VP1 108-116 (SEQ ID NO: 2136) A*02: 01 CLLPKMDSV large Tantigen 398-406 (SEQ ID NO: 2137) A*02: 01 FLWGPRALV MAGEA3 271-279 (SEQ ID NO: 2138) A*02: 01 IMDQVPFSV gp100 (pmel17) 209-217 (SEQ ID NO: 2139) A*02: 01 YLEPGPVTV gp100 (pmel) 280-288 (288V) (SEQ ID NO: 2140) A*02: 01 YLSGADLNL Carcinoembryonic antigen (CEA)-derived (SEQ ID NO: 2141) peptide CAP1-6D A*02: 01 SLLMWITQC NY-ESO-1 157-165 (9C) (SEQ ID NO: 2142) A*02: 01 KTWGQYWQV gp100 (pmel17) 154-162 (SEQ ID NO: 2143) A*02: 01 YLEPGPVTA gp100 (SEQ ID NO: 2144) A*02: 01 YMDGTMSQV Tyrosinase 369-377 (371D) (SEQ ID NO: 2145) A*02: 01 YLSGANLNL Carcinogenic Embryonic Antigen (CEA) (SEQ ID NO: 2146) 571-579 A*02: 01 ELAGIGILTV MelanA/MART26-35 (SEQ ID NO: 2147) A*02: 01 ILAKFLHWL Telomerase 540-548 (SEQ ID NO: 2148) A*02: 01 ALQPGTALL Prostate Stem Cell Antigen (SEQ ID NO: 2149) (PSCA) 14-22 A*02: 01 VISNDVCAQV Prostate Specific Antigen-1 (SEQ ID NO: 2150) (PSA-1) 154-163 A*02: 01 RLVDDFLLV Telomerase Reverse (SEQ ID NO: 2151) Transcriptase 865-873 A*02: 01 GVLVGVALI Carcinogenic Embryonic Antigen (SEQ ID NO: 2152) (CEA) 694-702 A*02: 01 VLYRYGSFSV gp100 (pmel17) 476-485 (SEQ ID NO: 2153) A*02: 01 PLFQVPEPV Alpha-fetoprotein isoform 1 137-145 (SEQ ID NO: 2154) A*02: 01 FMNKFIYEI Human alfa fetoprotein 158-166 (SEQ ID NO: 2155) A*02: 01 GLSPNLNRFL Alpha-fetoprotein isoform 2 167-176 (SEQ ID NO: 2156) A*02: 01 KVLEYVIKV MAGEA1 278-286 (SEQ ID NO: 2157) A*02: 01 LLGRNSFEV p53 264-272 (SEQ ID NO: 2158) A*02: 01 LLLLTVLTV MUC-1 12-20 (SEQ ID NO: 2159) A*02: 01 ILHNGAYSL HER-2/neu 435-443 (SEQ ID NO: 2160) A*02: 01 RLLQETELV HER-2/neu 689-697 (SEQ ID NO: 2161) A*02: 01 KIFGSLAFL HER-2/neu 369-377 (SEQ ID NO: 2162) A*02: 01 LLLLDVAPL HSP1A 459-467 (SEQ ID NO: 2163) A*02: 01 LLDVAPLSL HSP1A 461-469 (SEQ ID NO: 2164) A*02: 01 HLYQGCQVV Receptor tyrosine-protein kinase (SEQ ID NO: 2165) erbB-2 48-56 A*02: 01 HLSTAFARV G250 (renal cell carcinoma) 217-225 (SEQ ID NO: 2166) A*02: 01 VLQELNVTV Leukocyte Proteinase-3 (SEQ ID NO: 2167) (Wegener's autoantigen) 169-177 A*02: 01 KVAELVHFL MAGEA3 112-120 (SEQ ID NO: 2168) A*02: 01 VLAGVGFFI EPHA2 550-558 (SEQ ID NO: 2169) A*02: 01 FLYTLLREV STEAP 86-94 (SEQ ID NO: 2170) A*02: 01 ILLWQPIPV Prostatic Acid Phosphatase-3 (PAP-3) (SEQ ID NO: 2171) 135-143 A*02: 01 RLQEERTCKV BIR (SEQ ID NO: 2172) A*02: 01 QLCPICRAPV Livin/ML-IAP280 175-184 (SEQ ID NO: 2173) A*02: 01 VLGEAWRDQV TRAP 45-54 (SEQ ID NO: 2174) A*02: 01 LLLTVLTVV Tumor Mucin Antigen 13-21 (SEQ ID NO: 2175) A*02: 01 GLYDGMEHL MAGEA-10 254-262 (SEQ ID NO: 2176) A*02: 01 SLLMWITQV NY-ESO-1 157-165 (SEQ ID NO: 2177) A*02: 01 LMLGEFLKL Survivin 96-104 (SEQ ID NO: 2178) A*02: 01 YLFFYRKSV mTERT 572-580 (SEQ ID NO: 2179) A*02: 01 ELTLGEFLKL survivin 95-104 (SEQ ID NO: 2180) A*02: 01 FLTPKKLQCV Prostate Specific Antigen-1 (PSA-1) (SEQ ID NO: 2181) 141-150 A*02: 01 KLQCVDLHV Prostate Specific Antigen 146-154 (SEQ ID NO: 2182) A*02: 01 TLAPATEPA Mucin 79-87 (SEQ ID NO: 2183) A*02: 01 YLQVNSLQTV Telomerase Reverse Transcriptase (SEQ ID NO: 2184) (hTRT) 988-997 A*02: 01 SLGEQQYSV WT1 187-195 (SEQ ID NO: 2185) A*02: 01 SLEENIVIL RHAMM 275-283 (SEQ ID NO: 2186) A*02: 01 YMNGTMSQV Tyrosinase 368-376 (SEQ ID NO: 2187) A*02: 01 ILSLELMKL Receptor for hyaluronic acid- (SEQ ID NO: 2188) mediatedmotility (RHAMM) 165-173 A*02: 01 PLFDFSWLSL Bcl-2 208-217 (SEQ ID NO: 2189) A*02: 01 LLGATCMFV CyclinD 101-109 (SEQ ID NO: 2190) A*02: 01 ALYVDSLFFL PRAME PRA 300-309 (SEQ ID NO: 2191) A*02: 01 GLMEEMSAL Human Mena protein (overexpressed in (SEQ ID NO: 2192) breast cancer) A*02: 01 TMNGSKSPV hMena 502-510 (SEQ ID NO: 2193) A*02: 01 GVYDGREHTV MAGE-A4 230-239 (SEQ ID NO: 2194) A*02: 01 YLNDHLEPWI Bcl-X 173-182 (SEQ ID NO: 2195) A*02: 01 ALDVYNGLL Prostatic acid phosphatase precursor (SEQ ID NO: 2196) (PAP) 299-307 A*02: 01 ALFDIESKV PSM P2 (prostate) (SEQ ID NO: 2197) A*02: 01 SLAMLDLLHV Mutant anaplastic lymphoma kinase (SEQ ID NO: 2198) 1220-1229 A*02: 01 YLNTVQPTCV EGF-R 1138-1147 (SEQ ID NO: 2199) A*02: 01 KLFGTSGQKT EGF-R-479 350-359 (SEQ ID NO: 2200) A*02: 01 RMPEAAPPV p53 65-73 (SEQ ID NO: 2201) A*02: 01 PLTSIISAV Receptor tyrosine-protein kinase erb6-2 (SEQ ID NO: 2202) 728-736 A*02: 01 VLAGGFFLL PSMA 27-38 (SEQ ID NO: 2203) A*02: 01 LLHETDSAV PSMA/PSM-P1 4-12 (SEQ ID NO: 2204) A*02: 01 VMAGVGSPYV Receptor tyrosine-protein kinase erb6-2 (SEQ ID NO: 2205) 819-828 A*02: 01 VLPLTVAEV Mesothelin 530-538 (SEQ ID NO: 2206) A*02: 01 SLLFLLFSL Mesothelin 20-28 (SEQ ID NO: 2207) A*02: 01 QLFEELQEL Heme oxygenase-1 212-220 (SEQ ID NO: 2208) A*02: 01 VLDGLDVLL PRAME 100-108 (SEQ ID NO: 2209) A*02: 01 RLASFYDWPL BIR7 90-99 (SEQ ID NO: 2210) A*02: 01 LIAHNQVRQV HER-2/neu (85-94) (SEQ ID NO: 2211) A*02: 01 ILHDGAYSL HER-2 434-443 (SEQ ID NO: 2212) A*02: 01 FVGEFFTDV GPC3 144-152 (overexpressed in (SEQ ID NO: 2213) hepatocellular carcinoma) A*02: 01 LLLIWFRPV BKV Ltag 579-587 (SEQ ID NO: 2214) A*02: 01 KLQDASAEV HM1.24-aa 126-134 (SEQ ID NO: 2215) A*02: 01 SLYSFPEPEA PRAME (SEQ ID NO: 2216) A*02: 01 SLLQHLIGL PRAME 425-433 (SEQ ID NO: 2217) A*02: 01 VIFDFLHCI BKV Ltag 406-414 (SEQ ID NO: 2218) A*02: 01 VLDFAPPGA WT1 (SEQ ID NO: 2219) A*02: 01 TLPGYPPHV PAX-5 311-319 (SEQ ID NO: 2220) A*02: 01 YMEHNNVYTV Fibromodulin 250-259 (SEQ ID NO: 2221) A*02: 01 YLQHNEIQEV Fibromodulin 206-215 (SEQ ID NO: 2222) A*02: 01 SLVDVMPWL Cytochrome p450 161 239-248 (SEQ ID NO: 2223) A*02: 01 RLMNDMTAV HSP105 128-136 (SEQ ID NO: 2224) A*02: 01 RLARLALVL Trophoblast glycoprotein 17-25 (SEQ ID NO: 2225) A*02: 01 FLTGNQLAV 5T4 97-105 (SEQ ID NO: 2226) A*02: 01 LLLAGLFSL Fibromodulin 7-15 (SEQ ID NO: 2227) A*02: 01 FLGYLILGV Prostatic Acid Phosphatase-3 (PAP-3) (SEQ ID NO: 2228) A*02: 01 SLFLGILSV CD20 188-196 (B cell malignancies) (SEQ ID NO: 2229) A*02: 01 AVLPLLELV MCL-1 139-147 (SEQ ID NO: 2230) A*02: 01 SLSEKTVLL CD59 glycoprotein precursor 106-114 (SEQ ID NO: 2231) A*02: 01 YMCSFLFNL Ewing Tumor EZH2 666-674 (SEQ ID NO: 2232) A*02: 01 YLISGDSPV CD33 65-73 (1Y2L) (SEQ ID NO: 2233) A*02: 01 KASEKIFYV SSX2 41-49 (SEQ ID NO: 2234) A*02: 01 FLAKLNNTV HCA587 317-325 (SEQ ID NO: 2235) A*02: 01 GLAPPQHLIRV p53 187-197 (SEQ ID NO: 2236) A*02: 01 VIM PCSWWV Chondromodulin-I 319-327 (SEQ ID NO: 2237) A*02: 01 KVVEFLAML MAGE-C1 1083-1091 (SEQ ID NO: 2238) A*02: 01 LTLGEFLKL Survivin-3A 96-104 (SEQ ID NO: 2239) A*02: 01 ALPFGFILV IL13R 345-353 (SEQ ID NO: 2240) A*02: 01 TLADFDPRV EphA2 (SEQ ID NO: 2241) A*02: 01 ALMEQQHYV ITGB8 662-670 (SEQ ID NO: 2242) A*02: 01 CLTSTVQLV HER-2/neu 789-797 (SEQ ID NO: 2243) A*02: 01 GLLGASVLGL Telomerase Reverse Transcriptase (hTRT) (SEQ ID NO: 2244) 674-683 A*02: 01 QLLDGFMITL PASD1 39-48 (SEQ ID NO: 2245) A*02: 01 YLVGNVCIL PASD1 168-176 (SEQ ID NO: 2246) A*02: 01 ALLTSRLRFI Telomerase Reverse Transcriptase (SEQ ID NO: 2247) (hTRT) 615-624 A*02: 01 RLSSCVPVA TGF beta receptor type-2 131-139 (SEQ ID NO: 2248) A*02: 01 FLYDDNQRV Topoisomerase II-alpha-b 828-836 (SEQ ID NO: 2249) A*02: 01 YLIELIDRV TACE 250-258 (SEQ ID NO: 2250) A*02: 01 FLAEDALNTV Epithelial Discoidin Domain Receptor 1 (SEQ ID NO: 2251) (EDDR1) 867-876 A*02: 01 GLMKYIGEV TRPM8 187-195 (SEQ ID NO: 2252) A*02: 01 AILALLPAL Prostate Stem Cell Antigen (PSCA) (SEQ ID NO: 2253) 105-133 A*02: 01 GLQHWVPEL BA46 (Lactadherin) 97-106 (SEQ ID NO: 2254) A*02: 01 GVRGRVEEI BCR-ABL (SEQ ID NO: 2255) A*02: 01 ITDQVPFSV gp100 (pmel) 209-217 (SEQ ID NO: 2256) A*02: 01 KLCPVQLWV p53 139-147 (SEQ ID NO: 2257) A*02: 01 KVAEELVHFL MAGEA3 112-120 (alternative version) (SEQ ID NO: 2258) A*02: 01 SLPPPGTRV p53 149-157 (SEQ ID NO: 2259) A*02: 01 YLGSYGFRL p53 103-111 (SEQ ID NO: 2260) A*02: 01 YLQLVFGIEV MAGEA2 157-166 (SEQ ID NO: 2261) A*02: 01 TLQDIVYKL BMI1 74-82 (SEQ ID NO: 2262) A*02: 01 YAIDLPVSV L-dopachrome tautomerase 488-496 (SEQ ID NO: 2263) A*02: 01 AMVGAVLTA Tyrosinase 482-190 (SEQ ID NO: 2264) A*02: 01 ATVGIMIGV CEACAM5 687-695 (SEQ ID NO: 2265) A*02: 01 YVDPVITSI Hepatocyte growth factor receptor (SEQ ID NO: 2266) 673-681 A*02: 01 GVLLWEIFSL VEGFR1 28-37 (SEQ ID NO: 2267) A*02: 01 LMAQEALAFL CAMEL 2-11 (SEQ ID NO: 2268) A*02: 01 RVA(PHOSPHO-S)PTSGV Insulin receptor substrate-2 1097-1105 (SEQ ID NO: 2269) A*02: 01 RVASPTSGV IRS-2 1097-1105 (SEQ ID NO: 2270) A*02: 01 ALNVYNGLL ACPP 299-307 (SEQ ID NO: 2271) A*02: 01 ALSPVPPVV Bcl-2 85-93 (SEQ ID NO: 2272) A*02: 01 ALVCYGPGI FAP alpha 463-471 (SEQ ID NO: 2273) A*02: 01 ALWPWLLMAT RNF43 11-20 (SEQ ID NO: 2274) A*02: 01 ALYLMELTM CB9L2 (SEQ ID NO: 2275) A*02: 01 CLPSPSTPV BMI1 271-279 (SEQ ID NO: 2276) A*02: 01 ELSDSLGPV PASD1 695-703 (SEQ ID NO: 2277) A*02: 01 FLFLRNFSL TARP(V28L)27-35 (SEQ ID NO: 2278) A*02: 01 FLPSPLFFFL TARP(P5L) 5-13 (SEQ ID NO: 2279) A*02: 01 GLFKCGIAV FAP 639-647 (SEQ ID NO: 2280) A*02: 01 GLIQLVEGV TRAG-3 4-12 (SEQ ID NO: 2281) A*02: 01 ILGVLTSLV DLK1 309-317 (SEQ ID NO: 2282) A*02: 01 LLVPTCVFLV 691-700 (SEQ ID NO: 2283) A*02: 01 MLAVFLPIV STEAP 292-300 (293L) (SEQ ID NO: 2284) A*02: 01 NLFETPVEA 194-202 (SEQ ID NO: 2285) A*02: 01 QLGEQCWTV PSCA 44-51 (51A) (SEQ ID NO: 2286) A*02: 01 RLAEYQAYI SART3 309-317 (SEQ ID NO: 2287) A*02: 01 SIDWFMVTV p31-39 (SEQ ID NO: 2288) A*02: 01 SILLRDAGLV TRAG-3 57-66 (SEQ ID NO: 2289) A*02: 01 SLFEPPPPG PSMA 85-93 (SEQ ID NO: 2290) A*02: 01 SQADALKYV EZH2 729-737 (SEQ ID NO: 2291) A*02: 01 WLSLKTLLSL Bcl-2 214-223 (SEQ ID NO: 2292) A*02: 01 YLNRHLHTWI BCL-2 180-189 (SEQ ID NO: 2293) A*02: 01 YLQWIEFSI Prominin1 744-752 (SEQ ID NO: 2294) A*02: 01 YLYQWLGAPV Osteocalcin 51-60 (SEQ ID NO: 2295) A*02: 01 KLMSSNSTDL HSP105 234-243 (SEQ ID NO: 2296) A*02: 01 RLQGISPKI SSX2 103-111 (SEQ ID NO: 2297) A*02: 01 AILALLPALL PSCA (SEQ ID NO: 2298) A*02: 01 ALIHHNTHL HER2 466-474 (SEQ ID NO: 2299) A*02: 01 CMHLLLEAV MG50 624-632 (SEQ ID NO: 2300) A*02: 01 FLIIWQNTM F5P26 (SEQ ID NO: 2301) A*02: 01 FLPWHRLFLL Tyrosinase 207-216 (SEQ ID NO: 2302) A*02: 01 FVWLHYYSV TRP2 185-193(L) (SEQ ID NO: 2303) A*02: 01 GLFGDIYLA CSNK1A1 26-34 (SEQ ID NO: 2304) A*02: 01 GLFGDIYLAI CSNK1A1 26-35 (SEQ ID NO: 2305) A*02: 01 ILLRDAGLV TRAG-3L 58-66 (SEQ ID NO: 2306) A*02: 01 ILLVVVLGV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2307) 707-715 A*02: 01 ILNAMIAKI HAUS3 154-162 (SEQ ID NO: 2308) A*02: 01 KASEYLQLV MAGEA2 153-161 (SEQ ID NO: 2309) A*02: 01 KIWEELSVL MAGEA3 220-228 (SEQ ID NO: 2310) A*02: 01 KLIDRTE(S)L LSP1 325-333 (SEQ ID NO: 2311) A*02: 01 KLTGDENFTI Tyrosinase precursor 224-233 (SEQ ID NO: 2312) A*02: 01 LLCYSCKAQV PSCA 17-26 (SEQ ID NO: 2313) A*02: 01 LLLEAVPAV MG50 69-77 (SEQ ID NO: 2314) A*02: 01 LLNQLQVNL Mucin2 467-475 (SEQ ID NO: 2315) A*02: 01 LLRDAGLVKM TRAP 59-68 (SEQ ID NO: 2316) A*02: 01 LLRRYNVAKV SOX11 266-275 (SEQ ID NO: 2317) A*02: 01 LLSHGAVIEV Ankyrin NYBR1 158-167 (SEQ ID NO: 2318) A*02: 01 LVFGIELMEV MAGEA3 160-169 (SEQ ID NO: 2319) A*02: 01 LVFGIEVVEV MAGEA12 160-169 (SEQ ID NO: 2320) A*02: 01 MLWGWREHV Mucin2 645-653 (SEQ ID NO: 2321) A*02: 01 PLQPEQLQV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2322) 437-445 A*02: 01 QLMAFNHLI PAX3/FKHR 135-143 (SEQ ID NO: 2323) A*02: 01 QLMPYGCLL Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2324) 845-853 A*02: 01 RLGPTLMCL MG50 1244-1252 (SEQ ID NO: 2325) A*02: 01 RLTRFLSRV CyclinD 228-236 (SEQ ID NO: 2326) A*02: 01 RTF(S)PTYGL Desmuslin 426-434 (SEQ ID NO: 2327) A*02: 01 SILLRDAGL TRAP 57-65 (SEQ ID NO: 2328) A*02: 01 SLADEAEVYL GAS7 Neoepitope (SEQ ID NO: 2329) A*02: 01 SLDDYNHLV L-dopachrome tautomerase 288-296 (SEQ ID NO: 2330) A*02: 01 SLYKFSPFPL O-linked N-acetylglucosamine (SEQ ID NO: 2331) transferase FSPO6 A*02: 01 SMTR(S)PPRV SFRS2B 241-249 (SEQ ID NO: 2332) A*02: 01 TLEEITGYL Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2333) 448-456 A*02: 01 TLHCDCEIL MG50 210-218 (SEQ ID NO: 2334) A*02: 01 VLEPPGARDV BIR 7 230-239 (SEQ ID NO: 2335) A*02: 01 VLLALLMAGL Prostate stem cell antigen 4-13 (SEQ ID NO: 2336) A*02: 01 VLSVNVPDV MG50 625-633 (SEQ ID NO: 2337) A*02: 01 VLVKSPNHV Receptor tyrosine-protein kinase erbB-4 (SEQ ID NO: 2338) 890-898 A*02: 01 VMIG(S)PKKV Tensin3 1558-1566 (SEQ ID NO: 2339) A*02: 01 VVLGVVFGI Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2340) 743-751 A*02: 01 WLPKILGEV MG50 1051-1059 (SEQ ID NO: 2341) A*02: 01 WLQYFPNPV Cytochrome P450 246-254 (SEQ ID NO: 2342) A*02: 01 YLLDLSTNHL Fibromodulin 7-15 (SEQ ID NO: 2343) A*02: 01 YLWWVNNQSL CEA 176-185 (SEQ ID NO: 2344) A*02: 01 ALGGHPLLGV Dickkopf-related protein 1 20-29 (SEQ ID NO: 2345) A*02: 01 ALLAGLVSL FGFR4 676-684 (SEQ ID NO: 2346) A*02: 01 ALLTYMIAHI Thymidylate synthase 231-240 (SEQ ID NO: 2347) A*02: 01 ALMDKSLHV MART-1 56-64 (SEQ ID NO: 2348) A*02: 01 ALPPPLMLL Heparanase 8-16 (SEQ ID NO: 2349) A*02: 01 ALSVMGVYV MAGEA9 223-231 (SEQ ID NO: 2350) A*02: 01 ALVEFEDVL hnRNP L 140-148 (SEQ ID NO: 2351) A*02: 01 ALWPWLLMA RNF43 11-19 (SEQ ID NO: 2352) A*02: 01 AMLGTHTMEV Melanocyte-specific secreted (SEQ ID NO: 2353) glycoprotein 184-193 A*02: 01 AVIGALLAV Melanocyte-specific secreted (SEQ ID NO: 2354) glycoprotein 20-28 A*02: 01 CLYGNVEKV hnRNP L 404-412 (SEQ ID NO: 2355) A*02: 01 DLIFGLNAL Heparanase 185-193 (SEQ ID NO: 2356) A*02: 01 ELFQDLSQL ETV5 54-53 (SEQ ID NO: 2357) A*02: 01 FAWERVRGL Cyclin-dependent kinase inhibitor 1 (SEQ ID NO: 2358) 97-105 A*02: 01 FIASNGVKLV ACTN4 118-127 (K5N) (SEQ ID NO: 2359) A*02: 01 FLALIICNA Tubulin beta 4 283-291 (SEQ ID NO: 2360) A*02: 01 FLDEFMEGV Malic enzyme 224-232 (SEQ ID NO: 2361) A*02: 01 RMFPNAPYL WT-1 126-134 (Wilms tumor) (SEQ ID NO: 2362) A*02: 01 RLNMFTPYI Chlamydia trachomatis MOMP 258-266 (SEQ ID NO: 2363) A*02: 01 NMFTPYIGV MOMP precursor 283-291 (SEQ ID NO: 2364) A*02: 01 NLVPMVATV HCMV pp65 495-504 (SEQ ID NO: 2365) A*02: 01 VLEETSVML HCMV IE1 316-324 (UL123) (SEQ ID NO: 2366) A*02: 01 VLAELVKQI HCMV IE1 81-89 (SEQ ID NO: 2367) A*02: 01 MLNIPSINV pp65 120-128 (SEQ ID NO: 2368) A*02: 01 LLLDRLNQL SARS-CoV Nucleocapsid protein 223-231 (SEQ ID NO: 2369) (conserved in SARS-CoV-2) A*02: 01 FIAGLIAIV SARS-CoV-2 Spike glycoprotein 1220-1228 (SEQ ID NO: 2370) (confirmed epitope) A*02: 01 ALNTLVKQL SARS-CoV Spike glycoprotein precursor (SEQ ID NO: 2371) 940-948 (conserved in SARS-CoV-2) A*02: 01 LITGRLQSL SARS-CoV-2 Spike glycoprotein 996-1004 (SEQ ID NO: 2372) (confirmed epitope) A*02: 01 NLNESLIDL SARS-CoV (SEQ ID NO: 2373) Spike glycoprotein precursor 1174-1182 (conserved in SARS-Cov-2) A*02: 01 VLNDILSRL SARS-CoV Spike glycoprotein precursor (SEQ ID NO: 2374) 958-966 (conserved in SARS-Cov-2) A*02: 01 YLQPRTFLL SARS-Cov-2 Spike glycoprotein 269-277 (SEQ ID NO: 2375) (confirmed epitope) A*02: 01 LLYDANYFL SARS-CoV-2 ORF3a 139-147 (confirmed (SEQ ID NO: 2376) epitope) A*02: 01 RLQSLQTYV SARS-CoV-2 Spike glycoprotein 1000-1008 (SEQ ID NO: 2377) (confirmed subdominant epitope) A*02: 01 KLWAQCVQL SARS-CoV-2 ORF1ab 3886-3894 (confirmed (SEQ ID NO: 2378) epitope) A*02: 01 TLYAVATTI Dengue NS4b 40-48 (SEQ ID NO: 2379) A*02: 01 KLAEAIFKL Dengue NS5 563-571 (SEQ ID NO: 2380) A*02: 01 ILIRTGLLVI Dengue NS2b 97-106 (SEQ ID NO: 2381) A*02: 01 AIKRGLRTL Dengue NS3 112-120 (SEQ ID NO: 2382) A*02: 01 LLLGLMILL Dengue NS4a 56-64 (SEQ ID NO: 2383) A*02: 01 VLLLVTHYA Dengue NS4b 111-119 (SEQ ID NO: 2384) A*02: 01 GLCTLVAML EBV BMLF-1 259-267 (SEQ ID NO: 2385) A*02: 01 CLGGLLTMV EBV LMP-2 426-434 (SEQ ID NO: 2386) A*02: 01 YLLEMLWRL EBV LMP-1 125-133 (SEQ ID NO: 2387) A*02: 01 YLQQNWWTL EBV LMP1 159-167 (SEQ ID NO: 2388) A*02: 01 YVLDHLIVV EBV BRLF1 109-117 (SEQ ID NO: 2389) A*02: 01 FLYALALLL EBV LMP-2 356-364 (SEQ ID NO: 2390) A*02: 01 TLDYKPLSV EBV BMRF1 208-216 (SEQ ID NO: 2391) A*02: 01 LLDFVRFMGV EBV EBNA-3C 284-293 (SEQ ID NO: 2392) A*02: 01 FLDKGTYTL EBV BALF-4 276-284 (SEQ ID NO: 2393) A*02: 01 FLPSDFFPSV HBV core antigen 18-27 (SEQ ID NO: 2394) A*02: 01 FLLTRILTI HBV envelope 183-191 (SEQ ID NO: 2395) A*02: 01 GLSPTVWLSV HBV surface antigen 185-194 (SEQ ID NO: 2396) A*02: 01 WLSLLVPFV HBV surface antigen 172-181 (SEQ ID NO: 2397) A*02: 01 FLLSLGIHL HBV polymerase 573-581 (SEQ ID NO: 2398) A*02: 01 FLPSDFFPSI HBV core 18-27 (subtype ADR4) (SEQ ID NO: 2399) A*02: 01 VLHKRTLGL HBV X 92-100 (SEQ ID NO: 2400) A*02: 01 GLSRYVARL HBV Pol 455-463 (SEQ ID NO: 2401) A*02: 01 YMDDVVLGA HBV Polymerase 548-556 (SEQ ID NO: 2402) A*02: 01 KLHLYSHPI HBV Pol 502-510 (SEQ ID NO: 2403) A*02: 01 ELMTLATWV HBV core protein 64-72 (SEQ ID NO: 2404) A*02: 01 DLMGYIPAV HCV core 132-140 (SEQ ID NO: 2405) A*02: 01 CINGVCWTV HCV NS3 1073-1081 (SEQ ID NO: 2406) A*02: 01 YLLPRRGPRL HCV core 35-44 (SEQ ID NO: 2407) A*02: 01 VLSDFKTWL HCV NS5a 1987-1995 (SEQ ID NO: 2408) A*02: 01 ALYDVVTKL HCV NS5b 2594-2602 (SEQ ID NO: 2409) A*02: 01 KLVALGINAV HCV NS3 1406-1415 (SEQ ID NO: 2410) A*02: 01 LLFNILGGWV HCV NS4b 1807-1816 (SEQ ID NO: 2411) A*02: 01 KLSGLGINAV HCV NS3 1406-1415 (SEQ ID NO: 2412) A*02: 01 DLMGYIPLV HCV core 132-140 (SEQ ID NO: 2413) A*02: 01 CVNGVCWTV HCV NS3 1073-1081 (SEQ ID NO: 2414) A*02: 01 GLQDCTMLV HCV NS5B 2727-2735 (SEQ ID NO: 2415) A*02: 01 SLYNTVATL HIV-1 gag p17 76-84 (SEQ ID NO: 2416) A*02: 01 ILKEPVHGV HIV-1 RT476-484 (SEQ ID NO: 2417) A*02: 01 TLNAWVKVV HIV-1 gag p24 19-27 (SEQ ID NO: 2418) A*02: 01 KLTPLCVTL HIV-1 env gp120 90-98 (SEQ ID NO: 2419) A*02: 01 GLADQUHL HIV-1 vif 101-109 (SEQ ID NO: 2420) A*02: 01 LTFGWCFKL HIV-1 nef 137-145 (SEQ ID NO: 2421) A*02: 01 FLGKIWPS Gag 433-440 (SEQ ID NO: 2422) A*02: 01 ALVEMGHHA HIV Vpu 66-74 (SEQ ID NO: 2423) A*02: 01 RTLNAWVKV HIV gag 150-158 (SEQ ID NO: 2424) A*02: 01 NVWATHACV HIV env gp 67-7 (SEQ ID NO: 2425) A*02: 01 SLLNATAIAV HIV env 816-825 (SEQ ID NO: 2426) A*02: 01 SLFNTVATL HIV gag 77-85 (SEQ ID NO: 2427) A*02: 01 SLVKHHMYI HIV vif 23-31 (SEQ ID NO: 2428) A*02: 01 VIYHYVDDL HIV pol (SEQ ID NO: 2429) A*02: 01 YMLDLQPETT HPV 16 E7 11-20 (SEQ ID NO: 2430) A*02: 01 KLPQLCTEL HPV 16 E6 18-26 (SEQ ID NO: 2431) A*02: 01 YMLDLQPET HPV 16 E7 11-19 (SEQ ID NO: 2432) A*02: 01 MLDLQPETT HPV 16 E7 12-20 (SEQ ID NO: 2433) A*02: 01 VLMIKALEL Non muscle Myosin-9 741-749 (SEQ ID NO: 2434) A*02: 01 QLFNHTMFI Non-muscle Myosin 478-486 (SEQ ID NO: 2435) A*02: 01 QMARLAWEA 1116-1124 (SEQ ID NO: 2436) A*02: 01 LLFGYPVYV Human T-cell lymphotropic virus-1 (SEQ ID NO: 2437) (HTLV-1) tax 11-19 A*02: 01 AVLDGLLSL HTLV bZIP factor 42-50 (SEQ ID NO: 2438) A*02: 01 GLLSLEEEL bZIP factor 26-34 (SEQ ID NO: 2439) A*02: 01 GILGFVFTL Influenza A MP 58-66 (SEQ ID NO: 2440) A*02: 01 ILGFVFTLTV Influenza A MP 59-68 (SEQ ID NO: 2441) A*02: 01 KLGEFYNQMM Flu BNP 85-94 (Influenza B) (SEQ ID NO: 2442) A*02: 01 SITEVECFL VP1 36-44 (SEQ ID NO: 2443) A*02: 01 ILMWEAVTL VP1 100-108 (SEQ ID NO: 2444) A*02: 01 ALPHIIDEV LCMV envelope gp 10-18 (SEQ ID NO: 2445) A*02: 01 YLVSIFLHL LCMV envelope gp 447-455 (SEQ ID NO: 2446) A*02: 01 SLNQTVHSL NP 69-77 (SEQ ID NO: 2447) A*02: 01 YLNKIQNSL Plasmodium falciparum CSP 334-342 (SEQ ID NO: 2448) A*02: 01 FIDSYICQV miHAg H-Y (human SMCY) 311-319 (SEQ ID NO: 2449) A*02: 01 YIGEVLVSV HA-2 (SEQ ID NO: 2450) A*02: 01 VLHDDLLEA Minor Histocompatibility Antigen HA-1 (SEQ ID NO: 2451) 137-145 A*02: 01 RTLDKVLEV miHAg HA-8 (SEQ ID NO: 2452) A*02: 01 NEGATIVE Negative Control (SEQ ID NO: 2453) A*02: 01 TMFPHIIVDV Norovirus VP1 139-148 (SEQ ID NO: 2454) A*02: 01 LLDVPTAAV Interferon gamma inducible protein (SEQ ID NO: 2455) (GILT) 30 27-35 A*02: 01 RILGAVAKV Vinculin 822-830 (SEQ ID NO: 2456) A*02: 01 LMWYELSKI KSHVF-8 g6.492-500 (SEQ ID NO: 2457) A*02: 01 ILEDIVLTL Streptococcus pyogenes Cas9 615-623 (SEQ ID NO: 2458) A*02: 01 KMLKEMGEV RSV NP 137-145 (SEQ ID NO: 2459) A*02: 01 KLIANNTRV Mycobacterium bovis antigen 85-A (SEQ ID NO: 2460) 200-208 A*02: 01 GLPVEYLQV Mycobacterium bovis antigen 85-A 6-14 (SEQ ID NO: 2461) A*02: 01 GILTVSVAV 16 kDa (SEQ ID NO: 2462) A*02: 01 AMASTEGNV ESAT-6 (SEQ ID NO: 2463) A*02: 01 VLTDGNPPEV 19 kDa (SEQ ID NO: 2464) A*02: 01 KVDDTFYYV Vaccinia virus Host range protein (SEQ ID NO: 2465) 2 74-82 A*02: 01 ILDDNLYKV Vaccinia virus Copenhagen Protein (SEQ ID NO: 2466) G5 18-26 A*02: 01 ALWALPHAA 1E62 593-601 (SEQ ID NO: 2467) A*02: 01 RLDDDGNFQL West Nile Virus NY-99 polyprotein (SEQ ID NO: 2468) precursor (1452-1461) A*02: 01 ATWAENIQV West Nile virus NY-99 polyprotein (SEQ ID NO: 2469) precursor 3390-3398 A*02: 01 YTMDGEYRL West Nile virus NY-99 polyprotein (SEQ ID NO: 2470) precursor 2023-2031 A*02: 01 SVGGVFTSV WNV envelope gp 430-438 (SEQ ID NO: 2471) A*02: 01 SLFGQRIEV WNV nonstructural protein 4B 15-23 (SEQ ID NO: 2472) A*02: 01 LLWNGPMAV NS4B 214-222 (SEQ ID NO: 2473) A*03: 01 KQSSKALQR bcr-abl 210 kD fusion protein 21-29 (SEQ ID NO: 2474) A*03: 01 ALLAVGATK gp100 (pmel17) 17-25 (SEQ ID NO: 2475) A*03: 01 ATGFKQSSK bcr-abl 210 kD fusion protein 259-269 (SEQ ID NO: 2476) A*03: 01 RISTFKNWPK Survivin-3A 18-27 (27K) (SEQ ID NO: 2477) A*03: 01 RLGLQVRKNK RhoC 176-185 (1770 (SEQ ID NO: 2478) A*03: 01 RLLFFAPTR Mcl-1 95-103 (SEQ ID NO: 2479) A*03: 01 QVLKKIAQK HMOX1 145-153 (SEQ ID NO: 2480) A*03: 01 RIAAWMATY 165-173 (SEQ ID NO: 2481) A*03: 01 KLGGALQAK HCMV IE1 184-192 (SEQ ID NO: 2482) A*03: 01 KTFPPTEPK SARS-CoV-2 Nucleocapsid protein 362-370 (SEQ ID NO: 2483) (confirmed epitope) A*03: 01 ELERAADVK Dengue NS2b 52-60 (SEQ ID NO: 2484) A*03: 01 RVSTVQQLTK Dengue C 22-31 (SEQ ID NO: 2485) A*03: 01 RIEPSWADVK Dengue NS3 64-74 (SEQ ID NO: 2486) A*03: 01 RVIDPRRCMK Dengue NS3 422-431 (SEQ ID NO: 2487) A*03: 01 KITAEWLWK Dengue NS5 375-383 (SEQ ID NO: 2488) A*03: 01 RLRAEAQVK EBV EBNA 3A 603-611 (SEQ ID NO: 2489) A*03: 01 RVRAYTYSK EBV BRLF1 (SEQ ID NO: 2490) A*03: 01 RVCEKMALY HCV NS5B 2588-2596 (SEQ ID NO: 2491) A*03: 01 QVPLRPMTYK HIV-1 nef 73-82 (SEQ ID NO: 2492) A*03: 01 RLRPGGKKK HIV-1 gag p17 19-27 (SEQ ID NO: 2493) A*03: 01 AIFQSSMTK HIV pol 325-333 (SEQ ID NO: 2494) A*03: 01 KLCLRFLSK HPV 33 E6 64-72 (SEQ ID NO: 2495) A*03: 01 ILRGSVAHK Influenza A (PR8) NP 265-274 (SEQ ID NO: 2496) A*11: 01 KTFPPTEPK SARS-CoV-2 Nucleocapsid protein 362-370 (SEQ ID NO: 2483) (confirmed epitope) A*11: 01 GTSGSPIINR Dengue NS3 serotype 3&4 133-142 (SEQ ID NO: 2497) A*11: 01 GTSGSPIIDK Dengue NS3 133-142 (SEQ ID NO: 2498) A*11: 01 GTSGSPIVNR NS3 serotype 1 133-142 (SEQ ID NO: 2499) A*11: 01 GTSGSPIVDR Dengue NS3 serotype 2 133-142 (SEQ ID NO: 2500) A*11: 01 GTSGSPIADK Dengue NS3 133-142 (SEQ ID NO: 2501) A*11: 01 RVSTVQQLTK Dengue C 22-31 (SEQ ID NO: 2485) A*11: 01 RIEPSWADVK Dengue NS3 64-74 (SEQ ID NO: 2486) A*11: 01 RVIDPRRCMK Dengue NS3 422-431 (SEQ ID NO: 2487) A*11: 01 KITAEWLWK Dengue NS5 375-383 (SEQ ID NO: 2488) A*11: 01 IVTDFSVIK EBV EBNA-4 416-424 (SEQ ID NO: 2502) A*11: 01 SSCSSCPLSK EBV LMP-2 340-349 (SEQ ID NO: 2503) A*11: 01 ATIGTAMYK EBV BRLF1 134-142 (SEQ ID NO: 2504) A*11: 01 AVFDRKSDAK EBNA3B 399-408 (SEQ ID NO: 2505) A*11: 01 YVNVNMGLK HBV core antigen 88-96 (SEQ ID NO: 2506) A*11: 01 YVNTNMGLK HBV core 88-96 (SEQ ID NO: 2507) A*11: 01 STLPETTVVRR HBV core 141-151 (SEQ ID NO: 2508) A*11: 01 AVDLSHFLK HIV nef 84-92 (SEQ ID NO: 2509) A*11: 01 ACQGVGGPGHK HIV gag p24 (SEQ ID NO: 2510) A*11: 01 NTLEQTVKK HPV 33 E6 86-94 (SEQ ID NO: 2511) A*11: 01 SIIPSGPLK Influenza A MP 13-21 (SEQ ID NO: 2512) A*11: 01 RMVLASTTAK Influenza A MP1 178-187 (SEQ ID NO: 2513) A*11: 01 KSMREEYRK Influenza A MP2 70-78 (SEQ ID NO: 2514) A*24: 02 TYFSLNNKF Adenovirus 5 Hexon 37-45 (SEQ ID NO: 2515) A*24: 02 TYACFVSNL Carcinogenic Embryonic Antigen (SEQ ID NO: 2516) (CEA) 652-660 A*24: 02 AFLPWHRLF Tyrosinase 188-196 (SEQ ID NO: 2517) A*24: 02 IMPKAGLLI MAGE-A3 (SEQ ID NO: 2518) A*24: 02 VYFFLPDHL gp100-intron 4 (170-178) (SEQ ID NO: 2519) A*24: 02 EYLQLVFGI MAGEA2 156-164 (SEQ ID NO: 2520) A*24: 02 TYLPTNASL HER-2/neu 63-71 (SEQ ID NO: 2521) A*24: 02 VYGFVRACL Telomerase reverse transcriptase (SEQ ID NO: 2522) (hTRT) 461-469 A*24: 02 TFPDLESEF MAGEA3 97-105 (SEQ ID NO: 2523) A*24: 02 DYLQYVLQI MiHA ACC1 15-23 (SEQ ID NO: 2524) A*24: 02 RYCNLEGPPI Lymphocyte antigen 6 complex locus (SEQ ID NO: 2525) K (LY6K) 177-186 A*24: 02 AYACNTSTL Survivin 80-88 (SEQ ID NO: 2526) A*24: 02 CYASGWGSI Prostate Specific Antigen-1 153-161 (SEQ ID NO: 2527) A*24: 02 DYLNEWGSRF CDH3 807-816 (SEQ ID NO: 2528) A*24: 02 EYCPGGNLF MELK 87-95 (93N) (SEQ ID NO: 2529) A*24: 02 EYYELFVNI DEP DC1 294-302 (SEQ ID NO: 2530) A*24: 02 GYCTQIGIF HENMT1 221-229 (SEQ ID NO: 2531) A*24: 02 IYTWIEDHF FOXM1 262-270 (SEQ ID NO: 2532) A*24: 02 NYQPVWLCL RNF43 721-729 (722Y) (SEQ ID NO: 2533) A*24: 02 RYNAQCQETI Midkine 110-119 (SEQ ID NO: 2534) A*24: 02 EYRALQLHL CA9 219-227 (SEQ ID NO: 2535) A*24: 02 SYRNEIAYL TTK protein kinase 551-559 (SEQ ID NO: 2536) A*24: 02 VYLRVRPLL KIF20A 67-75 (SEQ ID NO: 2537) A*24: 02 VYYNWQYLL IL13r 146-154 (SEQ ID NO: 2538) A*24: 02 VYALPLKML HCMV pp65 113-121 (SEQ ID NO: 2539) A*24: 02 QYDPVAALF HCMV pp65 341-349 (SEQ ID NO: 2540) A*24: 02 AYAQKIFKI CMV 1E-1 248-256 (SEQ ID NO: 2541) A*24: 02 QYSDRRWCF Dengue NS3 557-565 (Singapore/S275/1990) (SEQ ID NO: 2542) A*24: 02 TYGPVFMCL EBV LMP-2 419-427 (SEQ ID NO: 2543) A*24: 02 PYLFWLAAI EBV LMP2 131-139 (SEQ ID NO: 2544) A*24: 02 TYGPVFMSL EBV LMP2 419-427 (SEQ ID NO: 2545) A*24: 02 EYLVSFGVW HBV core 117-125 (SEQ ID NO: 2546) A*24: 02 KYTSFPWLL HBV polymerase 756-764 (SEQ ID NO: 2547) A*24: 02 FFPSIRDLL HBV core protein 23-31 (SEQ ID NO: 2548) A*24: 02 AYSQQTRGL HCV NS3 1031-1039 (SEQ ID NO: 2549) A*24: 02 RYPLTFGWCY HIV-1 Nef 134-143 (SEQ ID NO: 2550) A*24: 02 RYLKDQQLL HIV-1 gag gp41 67-75 (SEQ ID NO: 2551) A*24: 02 RYLRDQQLL HIV env (SEQ ID NO: 2552) A*24: 02 RYPLTFGWCF HIV nef 143-152 (SEQ ID NO: 2553) A*24: 02 RYPLTFGW HIV nef (SEQ ID NO: 2554) A*24: 02 VYDFAFRDL HPV16 E6 (SEQ ID NO: 2555) A*24: 02 SFHSLHLLF HTLV Tax 301-309 (SEQ ID NO: 2556) A*29: 02 KEKYIDQEEL HSP90 alpha 280-288 (Pathologic (SEQ ID NO: 2557) Conditions) A*29: 02 LYNTVATLY HIV gag 79-86 (SEQ ID NO: 2558) A*29: 02 SFDPIPIHY HIV env 216-224 (SEQ ID NO: 2559) A*29: 02 SFNCRGEFFY HIV env 382-391 (SEQ ID NO: 2560) A*68: 01 TVSGNILTIR NY-ESO-1 127-136 (SEQ ID NO: 2561) B*07: 02 VPQYGYLTL AAV2 372-380 (SEQ ID NO: 2562) B*07: 02 KPYSGTAYNSL Adenovirus Hexon 114-124 (SEQ ID NO: 2563) B*07: 02 KPYSGTAYNAL Adenovirus Hexon 114-124 (SEQ ID NO: 2564) B*07: 02 LPLMRKAYL LT antigen 27-35 (SEQ ID NO: 2565) B*07: 02 LPWHRLFLL Tyrosinase 208-216 (SEQ ID NO: 2566) B*07: 02 EPR(PHOSPHO-S)PSHSM Insulin receptor substrate 2 (SEQ ID NO: 2567) B*07: 02 TPNQRQNVC P2X5 (SEQ ID NO: 2568) B*07: 02 APRGVRMAV LAGE-1 46-54 (SEQ ID NO: 2569) B*07: 02 LPVSPRLQL CEACAM 185-193 (SEQ ID NO: 2570) B*07: 02 TPRVTGGGAM HCMV pp65 417-426 (SEQ ID NO: 2571) B*07: 02 RPHERNGFTVL HCMV pp65 265-275 (SEQ ID NO: 2572) B*07: 02 SPRWYFYYL SARS-CoV-2 Nucleocapsid protein 105-113 (SEQ ID NO: 2573) (confirmed epitope) B*07: 02 APTRVVAAEM Dengue NS3 serotype 2 222-231 (SEQ ID NO: 2574) B*07: 02 RPPIFIRRL EBV EBNA-3A 247-255 (SEQ ID NO: 2575) B*07: 02 RPQGGSRPEFVKL EBV BMRF1 116-128 (SEQ ID NO: 2576) B*07: 02 QPRAPIRPI EBV EBNA-3C 881-889 (SEQ ID NO: 2577) B*07: 02 LPSDFFPSV HBV core 19-27 (SEQ ID NO: 2578) B*07: 02 GPRLGVRAT HCV core 41-49 (SEQ ID NO: 2579) B*07: 02 DPRRRSRNL HCV core 111-119 (SEQ ID NO: 2580) B*07: 02 IPRRIRQGL HIV-1 env gp120 848-856 (SEQ ID NO: 2581) B*07: 02 TPGPGVRYPL HIV-1 nef 128-137 (SEQ ID NO: 2582) B*07: 02 GPGHKARVL HIV gag p24 223-231 (SEQ ID NO: 2583) B*07: 02 KPTLKEYVL HPV 33 E7 5-13 (SEQ ID NO: 2584) B*07: 02 LPVSCPEDL bZIP factor 10-18 (SEQ ID NO: 2585) B*07: 02 QPEWFRNVL Influenza A PB1 329-337 (SEQ ID NO: 2586) B*07: 02 SPIVPSFDM Influenza A NP 473-481 (SEQ ID NO: 2587) B*07: 02 SPSVDKARAEL MiHAg SMCY 1041-1051 (SEQ ID NO: 2588) B*08: 01 GFKQSSKAL bcr-abl 210 kD fusion protein 19-27 (SEQ ID NO: 2589) B*08: 01 ELRRKMMYM 1E1 199-207 (SEQ ID NO: 2590) B*08: 01 ELKRKMIYM CMV 1E-1 199-207 (RA Position 3, M/I (SEQ ID NO: 2591) Position 7) B*08: 01 RIKQKGIL Dengue NS3 25-32 (SEQ ID NO: 2592) B*08: 01 LEKTKKDL Dengue NS4a 6-13 (SEQ ID NO: 2593) B*08: 01 FLRGRAYGL EBV EBNA-3A 193-201 (SEQ ID NO: 2594) B*08: 01 RAKFKQLL EBV BZLF-1 190-197 (SEQ ID NO: 2595) B*08: 01 QAKWRLQTL EBV EBNA3A 158-166 (SEQ ID NO: 2596) B*08: 01 GLKILQLL HBV external core Ag (SEQ ID NO: 2597) B*08: 01 HSKKKCDEL HCV NS3 1395-1403 (SEQ ID NO: 2598) B*08: 01 FLKEKGGL HIV-1 nef 90-97 (SEQ ID NO: 2599) B*08: 01 GEIYKRWII HIV-1 gag p24 261-269 (SEQ ID NO: 2600) B*08: 01 ElYKRWII HIV p24 gag 128-135 (SEQ ID NO: 2601) B*08: 01 YLKDQQLL Env 586-593 (SEQ ID NO: 2602) B*15: 01 CLIPTAMAF Dengue C 107-115 (SEQ ID NO: 2603) B*15: 01 RLRPGGKKKY HIV-1 p17 20-29 (SEQ ID NO: 2604) B*27: 05 GRFGLATEK BRAF594-601 (600E) (SEQ ID NO: 2605) B*27: 05 GRFGLATVK BRAF594-601 (600V) (SEQ ID NO: 2606) B*27: 05 RMFPNAPYL WT-1 126-134 (Wilms tumor) (SEQ ID NO: 2362) B*27: 05 ARKLLLDNL PqqC-like protein 70-78 (SEQ ID NO: 2607) B*27: 05 NRAKQVIKL Probable ATP-dependent Clp protease (SEQ ID NO: 2608) ATP-binding subunit 7-15 B*27: 05 QRNAPRITF SARS-CoV-2 Nucleocapsid protein 9-17 (SEQ ID NO: 2609) (confirmed epitope) B*27: 05 ARMILMTHF HCV NS5B 2841-2849 (SEQ ID NO: 2610) B*27: 05 KRWIILGLNK HIV-1 gag p24 265-274 (SEQ ID NO: 2611) B*27: 05 KRWIIMGLNK HIV-1 Gag p24 263-272 (SEQ ID NO: 2612) B*27: 05 GRAFVTIGK HIV-1 gp100 103-111 (SEQ ID NO: 2613) B*27: 05 SRYWAIRTR Influenza A NP 383-391 (SEQ ID NO: 2614) B*35: 01 IPYLDGTFY Adenovirus Hexon (SEQ ID NO: 2615) B*35: 01 MPFATPMEA NY-ESO-1 94-102 (SEQ ID NO: 2616) B*35: 01 IPSINVHHY HCMV pp65 123-131 (SEQ ID NO: 2617) B*35: 01 TPEGIIPTL Dengue NS3 500-508 (SEQ ID NO: 2618) B*35: 01 YPLHEQHGM EBV EBNA-3A 458-466 (SEQ ID NO: 2619) B*35: 01 EPLPQGQLTAY EBV BZLF-1 54-64 (SEQ ID NO: 2620) B*35: 01 HPVAEADYFEY EBV EBNA-1 407-417(4A) (SEQ ID NO: 2621) B*35: 01 HPNIEEVAL HCV NS3 1359-1367 (SEQ ID NO: 2622) B*35: 01 CPNSSIVY HCV E1 207-214 (SEQ ID NO: 2623) B*35: 01 NPDIVIYQY HIV-1 RT330-338 (SEQ ID NO: 2624) B*35: 01 NPDIVIYQY HIV-1 HIV-1 RT328-336 (SEQ ID NO: 2625) B*35: 01 VPLDEDFRKY HIV-1 HIV-1 RT273-282 (SEQ ID NO: 2626) B*40: 01 MEVTPSGTWL SARS-CoV-2 Nucleocapsid protein 322-330 (SEQ ID NO: 2627) (confirmed epitope) B*40: 01 GEARKTFVEL Dengue NS3 528-537 (SEQ ID NO: 2628) B*40: 01 IEDPPFNSL EBV LMP2 200-208 (SEQ ID NO: 2629) B*40: 01 REISVPAEIL HCV NS5a 2266-2275 (SEQ ID NO: 2630) B*40: 01 KEKGGLEGL HIV-1 Nef 92-100 (SEQ ID NO: 2631) C*06: 02 TRATKMQVI pp65 211-219 (SEQ ID NO: 2632) C*06: 02 QIKVRVDM 1E1 88-95 (SEQ ID NO: 2633) C*06: 02 TRRFLPQIL NS3 205-213 (SEQ ID NO: 2634) C*07: 02 CRVLCCYVL 1E1 309-317 (SEQ ID NO: 2635) E*01: 01 VMAPRTLIL HLA-C leader sequence peptide (SEQ ID NO: 2636) E*01: 01 VMAPRTLVL HLA-A leader sequence peptide (SEQ ID NO: 2637) A*01: 01 SADNNNSEY AAV VP1 492-500 (SEQ ID NO: 2082) A*01: 01 TDLGQNLLY Adenovirus 5 Hexon 886-894 (SEQ ID NO: 2083) A*01: 01 EADPTGHSY MAGE-A1 161-169 (SEQ ID NO: 2084) A*01: 01 EVDPIGHLY MAGE-A3 168-176 (SEQ ID NO: 2085) A*01: 01 KSDICTDEY Tyrosinase 243-251 (244S) (SEQ ID NO: 2086) A*01: 01 KCDICTDEY Tyrosinase 243-251 (SEQ ID NO: 2087) A*01: 01 QSLEIISRY Mcl-1 177-185 (SEQ ID NO: 2088) A*01: 01 YVDFREYEYY FLT3 ITD (SEQ ID NO: 2089) A*01: 01 TLDTLTAFY Mesothelin 429-437 (SEQ ID NO: 2090) A*01: 01 LTDDRLFTCY PLEKHM2 (SEQ ID NO: 2091) A*01: 01 DSDPDSFQDY Tyr Ala 454-463 (SEQ ID NO: 2092) A*01: 01 EADPIGHLY MAGEA3 (SEQ ID NO: 2093) A*01: 01 EVDPASNTY MAGE-A4 169-177 (SEQ ID NO: 2094) A*01: 01 HSTNGVTRIY PSMA (SEQ ID NO: 2095) A*01: 01 ILDTAGREEY N-ras 55-64 (SEQ ID NO: 2096) A*01: 01 LVDVMPWLQY Cytochrome P450 240-249 (SEQ ID NO: 2097) A*01: 01 RSDSGQQARY AIM-2 (SEQ ID NO: 2098) A*01: 01 VTEPGTAQY Minor antigen HA-3T (Lbc oncogene (SEQ ID NO: 2099) 451-459) A*01: 01 VYDFFVWLHY TRP-2 181-190 (SEQ ID NO: 2100) A*01: 01 YSEHPTFTSQY HCMV pp65 363-373 (SEQ ID NO: 2101) A*01: 01 VTEHDTLLY HCMV pp50 245-253 (SEQ ID NO: 2102) A*01: 01 FTSDYYQLY SARS-CoV-2 ORF3a 207-215 (SEQ ID NO: 2103) (confirmed epitope) A*01: 01 TTDPSFLGRY SARS-CoV-2 Replicase polyprotein lab (SEQ ID NO: 2638) 1637-1646 (confirmed epitope) A*01: 01 PTDNYITTY SARS-CoV-2 Replicase polyprotein lab (SEQ ID NO: 2639) 1621-1629 (confirmed epitope) A*01: 01 LLDTASALY HBV core 30-38 (SEQ ID NO: 2106) A*01: 01 ATDALMTGY HCV NS3 1435-1443 (SEQ ID NO: 2107) A*01: 01 ATDALMTGF HCV NS3 1436-1444 (SEQ ID NO: 2108) A*01: 01 CTELKLSDY Influenza A (PR8) NP 44-52 (SEQ ID NO: 2109) A*01: 01 VSDGGPNLY Influenza A PB1 591-599 (SEQ ID NO: 2110) A*01: 01 IVDCLTEMY DRRFY(1521-1529)) (SEQ ID NO: 2111) A*0201 ALCNTDSPL iLR1 (SEQ ID NO: 2112) A*0201 ALKDVEERV MAGE-C2 336-344 (SEQ ID NO: 2113) A*0201 LLAARAIVAI iLR1 59-68 (SEQ ID NO: 2114) A*0201 RLWQELSDI circadian clock protein PASD1 691-700 (SEQ ID NO: 2115) A*0201 LLFGLALIEV MAGE-C2 191-200 (SEQ ID NO: 2116) A*0201 FLDPRPLTV CYP190 (SEQ ID NO: 2117) A*0201 STLCQVEPV MPP11 (SEQ ID NO: 2118) A*0201 VLQMKEEDV iLR1 (SEQ ID NO: 2119) A*0201 AIQDLCLAV NPM1 (SEQ ID NO: 2120) A*0201 QLLIKAVNL MPP11 (SEQ ID NO: 2121) A*0201 AIQDLCVAV NPM1 (SEQ ID NO: 2122) A*0201 ALTPVVVTL cyclin-dependent kinase 4 170-178 (SEQ ID NO: 2123) A*02: 01 KLQVFLIVL T1D Diabetes human prepro islet amyloid (SEQ ID NO: 2640) polypeptide pplAPP 5-13 A*02: 01 VMNILLQYV GAD65 114-123 (SEQ ID NO: 2125) A*02: 01 SLSRFSWGA Myelin basic protein 110-118 (SEQ ID NO: 2126) A*02: 01 HLVEALYLV Insulin B chain 10-18 (SEQ ID NO: 2127) A*02: 01 LNIDLLWSV T1D Diabetes IGRP 228-236 (SEQ ID NO: 2128) A*02: 01 VLFGLGFAI T1D Diabetes IGRP 265-273 (SEQ ID NO: 2129) A*02: 01 ALWGPDPAAA Proinsulin precursor 15-24 (SEQ ID NO: 2130) A*02: 01 MVWESGCTV IA-2 797-805 (SEQ ID NO: 2131) A*02: 01 YTCPLCRAPV SSA SS-56 55-64 (SEQ ID NO: 2132) A*02: 01 VIVMLTPLV IA-2 805-813 (SEQ ID NO: 2133) A*02: 01 AITEVECFL VP1 44-52 (SEQ ID NO: 2134) A*02: 01 FLHCIVFNV large Tantigen 410-418 (SEQ ID NO: 2135) A*02: 01 LLMWEAVTV VP1 108-116 (SEQ ID NO: 2136) A*02: 01 CLLPKMDSV large Tantigen 398-406 (SEQ ID NO: 2137) A*02: 01 FLWGPRALV MAGEA3 271-279 (SEQ ID NO: 2138) A*02: 01 IMDQVPFSV gp100 (pmel17) 209-217 (SEQ ID NO: 2139) A*02: 01 YLEPGPVTV gp100 (pmel) 280-288 (288V) (SEQ ID NO: 2140) A*02: 01 YLSGADLNL Carcinoembryonic antigen (CEA)-derived (SEQ ID NO: 2141) peptide CAP1-6D A*02: 01 SLLMWITQC NY-ESO-1 157-165 (9C) (SEQ ID NO: 2142) A*02: 01 KTWGQYWQV gp100 (pmel17) 154-162 (SEQ ID NO: 2143) A*02: 01 YLEPGPVTA gp100 (SEQ ID NO: 2144) A*02: 01 YMDGTMSQV Tyrosinase 369-377 (371D) (SEQ ID NO: 2145) A*02: 01 YLSGANLNL Carcinogenic Embryonic Antigen (CEA) (SEQ ID NO: 2146) 571-579 A*02: 01 ELAGIGILTV MelanA/MART 26-35 (SEQ ID NO: 2147) A*02: 01 ILAKFLHWL Telomerase 540-548 (SEQ ID NO: 2148) A*02: 01 ALQPGTALL Prostate Stem Cell Antigen (PSCA) (SEQ ID NO: 2149) 14-22 A*02: 01 VISNDVCAQV Prostate Specific Antigen-1 (PSA-1) (SEQ ID NO: 2150) 154-163 A*02: 01 RLVDDFLLV Telomerase Reverse Transcriptase (SEQ ID NO: 2151) 865-873 A*02: 01 GVLVGVALI Carcinogenic Embryonic Antigen (CEA) (SEQ ID NO: 2152) 694-702 A*02: 01 VLYRYGSFSV gp100 (pmel17) 476-485 (SEQ ID NO: 2153) A*02: 01 PLFQVPEPV Alpha-fetoprotein isoform 1 137-145 (SEQ ID NO: 2154) A*02: 01 FMNKFIYEI Human alfa fetoprotein 158-166 (SEQ ID NO: 2155) A*02: 01 GLSPNLNRFL Alpha-fetoprotein isoform 2 167-176 (SEQ ID NO: 2156) A*02: 01 KVLEYVIKV MAGEA1 278-286 (SEQ ID NO: 2157) A*02: 01 LLGRNSFEV p53 264-272 (SEQ ID NO: 2158) A*02: 01 LLLLTVLTV MUC-1 12-20 (SEQ ID NO: 2159) A*02: 01 ILHNGAYSL HER-2/neu 435-443 (SEQ ID NO: 2160) A*02: 01 RLLQETELV HER-2/neu 689-697 (SEQ ID NO: 2161) A*02: 01 KIFGSLAFL HER-2/neu 369-377 (SEQ ID NO: 2162) A*02: 01 LLLLDVAPL HSP1A 459-467 (SEQ ID NO: 2163) A*02: 01 LLDVAPLSL HSP1A 461-469 (SEQ ID NO: 2164) A*02: 01 HLYQGCQVV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2165) 48-56 A*02: 01 HLSTAFARV G250 (renal cell carcinoma) 217-225 (SEQ ID NO: 2166) A*02: 01 VLQELNVTV Leukocyte Proteinase-3 (Wegener's (SEQ ID NO: 2167) autoantigen) 169-177 A*02: 01 KVAELVHFL MAGEA3 112-120 (SEQ ID NO: 2168) A*02: 01 VLAGVGFFI EPHA2 550-558 (SEQ ID NO: 2169) A*02: 01 FLYTLLREV STEAP 86-94 (SEQ ID NO: 2170) A*02: 01 ILLWQPIPV Prostatic Acid Phosphatase-3 (PAP-3) (SEQ ID NO: 2171) 135-143 A*02: 01 RLQEERTCKV BIR (SEQ ID NO: 2172) A*02: 01 QLCPICRAPV Livin/ML-IAP280 175-184 (SEQ ID NO: 2173) A*02: 01 VLGEAWRDQV TRAP 45-54 (SEQ ID NO: 2174) A*02: 01 LLLTVLTVV Tumor Mucin Antigen 13-21 (SEQ ID NO: 2175) A*02: 01 GLYDGMEHL MAGEA-10 254-262 (SEQ ID NO: 2176) A*02: 01 SLLMWITQV NY-ESO-1 157-165 (SEQ ID NO: 2177) A*02: 01 LMLGEFLKL Survivin 96-104 (SEQ ID NO: 2178) A*02: 01 YLFFYRKSV mTERT 572-580 (SEQ ID NO: 2179) A*02: 01 ELTLGEFLKL survivin 95-104 (SEQ ID NO: 2180) A*02: 01 FLTPKKLQCV Prostate Specific Antigen-1 (PSA-1) (SEQ ID NO: 2181) 141-150 A*02: 01 KLQCVDLHV Prostate Specific Antigen 146-154 (SEQ ID NO: 2182) A*02: 01 TLAPATEPA Mucin 79-87 (SEQ ID NO: 2183) A*02: 01 YLQVNSLQTV Telomerase Reverse Transcriptase (hTRT) (SEQ ID NO: 2184) 988-997 A*02: 01 SLGEQQYSV WT1 187-195 (SEQ ID NO: 2185) A*02: 01 SLEENIVIL RHAMM 275-283 (SEQ ID NO: 2186) A*02: 01 YMNGTMSQV Tyrosinase 368-376 (SEQ ID NO: 2187) A*02: 01 ILSLELMKL Receptor for hyaluronic acid-mediated (SEQ ID NO: 2188) motility (RHAMM) 165-173 A*02: 01 PLFDFSWLSL Bcl-2 208-217 (SEQ ID NO: 2189) A*02: 01 LLGATCMFV CyclinD 101-109 (SEQ ID NO: 2190) A*02: 01 ALYVDSLFFL PRAME PRA 300-309 (SEQ ID NO: 2191) A*02: 01 GLMEEMSAL Human Mena protein (overexpressed in (SEQ ID NO: 2192) breast cancer) A*02: 01 TMNGSKSPV hMena 502-510 (SEQ ID NO: 2193) A*02: 01 GVYDGREHTV MAGE-A4 230-239 (SEQ ID NO: 2194) A*02: 01 YLNDHLEPWI Bcl-X 173-182 (SEQ ID NO: 2195) A*02: 01 ALDVYNGLL Prostatic acid phosphatase precursor (SEQ ID NO: 2196) (PAP) 299-307 A*02: 01 ALFDIESKV PSM P2 (prostate) (SEQ ID NO: 2197) A*02: 01 SLAMLDLLHV Mutant anaplastic lymphoma kinase (SEQ ID NO: 2198) 1220-1229 A*02: 01 YLNTVQPTCV EGF-R 1138-1147 (SEQ ID NO: 2199) A*02: 01 KLFGTSGQKT EGF-R-479 350-359 (SEQ ID NO: 2200) A*02: 01 RMPEAAPPV p53 65-73 (SEQ ID NO: 2201) A*02: 01 PLTSIISAV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2202) 728-736 A*02: 01 VLAGGFFLL PSMA 27-38 (SEQ ID NO: 2203) A*02: 01 LLHETDSAV PSMA/PSM-P1 4-12 (SEQ ID NO: 2204) A*02: 01 VMAGVGSPYV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2205) 819-828 A*02: 01 VLPLTVAEV Mesothelin 530-538 (SEQ ID NO: 2206) A*02: 01 SLLFLLFSL Mesothelin 20-28 (SEQ ID NO: 2207) A*02: 01 QLFEELQEL Heme oxygenase-1 212-220 (SEQ ID NO: 2208) A*02: 01 VLDGLDVLL PRAME 100-108 (SEQ ID NO: 2209) A*02: 01 RLASFYDWPL BIR7 90-99 (SEQ ID NO: 2210) A*02: 01 LIAHNQVRQV HER-2/neu (85-94) (SEQ ID NO: 2211) A*02: 01 ILHDGAYSL HER-2 434-443 (SEQ ID NO: 2212) A*02: 01 FVGEFFTDV GPC3 144-152 (overexpressed in (SEQ ID NO: 2213) hepatocellular carcinoma) A*02: 01 LLLIWFRPV BKV Ltag 579-587 (SEQ ID NO: 2214) A*02: 01 KLQDASAEV HM1.24-aa 126-134 (SEQ ID NO: 2215) A*02: 01 SLYSFPEPEA PRAME (SEQ ID NO: 2216) A*02: 01 SLLQHLIGL PRAME 425-433 (SEQ ID NO: 2217) A*02: 01 VIFDFLHCI BKV Ltag 406-414 (SEQ ID NO: 2218) A*02: 01 VLDFAPPGA WT1 (SEQ ID NO: 2219) A*02: 01 TLPGYPPHV PAX-5 311-319 (SEQ ID NO: 2220) A*02: 01 YMEHNNVYTV Fibromodulin 250-259 (SEQ ID NO: 2221) A*02: 01 YLQHNEIQEV Fibromodulin 206-215 (SEQ ID NO: 2222) A*02: 01 SLVDVMPWL Cytochrome p450 161 239-248 (SEQ ID NO: 2223) A*02: 01 RLMNDMTAV HSP105 128-136 (SEQ ID NO: 2224) A*02: 01 RLARLALVL Trophoblast glycoprotein 17-25 (SEQ ID NO: 2225) A*02: 01 FLTGNQLAV 5T4 97-105 (SEQ ID NO: 2226) A*02: 01 LLLAGLFSL Fibromodulin 7-15 (SEQ ID NO: 2227) A*02: 01 FLGYLILGV Prostatic Acid Phosphatase-3 (PAP-3) (SEQ ID NO: 2228) A*02: 01 SLFLGILSV CD20 188-196 (B cell malignancies) (SEQ ID NO: 2229) A*02: 01 AVLPLLELV MCL-1 139-147 (SEQ ID NO: 2230) A*02: 01 SLSEKTVLL CD59 glycoprotein precursor 106-114 (SEQ ID NO: 2231) A*02: 01 YMCSFLFNL Ewing Tumor EZH2 666-674 (SEQ ID NO: 2232) A*02: 01 YLISGDSPV CD33 65-73 (1Y2L) (SEQ ID NO: 2233) A*02: 01 KASEKIFYV SSX2 41-49 (SEQ ID NO: 2234) A*02: 01 FLAKLNNTV HCA587 317-325 (SEQ ID NO: 2235) A*02: 01 GLAPPQHLIRV p53 187-197 (SEQ ID NO: 2236) A*02: 01 VIM PCSWWV Chondromodulin-I 319-327 (SEQ ID NO: 2237) A*02: 01 KVVEFLAML MAGE-C1 1083-1091 (SEQ ID NO: 2238) A*02: 01 LTLGEFLKL Survivin-3A 96-104 (SEQ ID NO: 2239) A*02: 01 ALPFGFILV IL13R 345-353 (SEQ ID NO: 2240) A*02: 01 TLADFDPRV EphA2 (SEQ ID NO: 2241) A*02: 01 ALMEQQHYV ITGB8 662-670 (SEQ ID NO: 2242) A*02: 01 CLTSTVQLV HER-2/neu 789-797 (SEQ ID NO: 2243) A*02: 01 GLLGASVLGL Telomerase Reverse Transcriptase (hTRT) (SEQ ID NO: 2244) 674-683 A*02: 01 QLLDGFMITL PASD1 39-48 (SEQ ID NO: 2245) A*02: 01 YLVGNVCIL PASD1 168-176 (SEQ ID NO: 2246) A*02: 01 ALLTSRLRFI Telomerase Reverse Transcriptase (hTRT) (SEQ ID NO: 2247) 615-624 A*02: 01 RLSSCVPVA TGF beta receptor type-2 131-139 (SEQ ID NO: 2248) A*02: 01 FLYDDNQRV Topoisomerase II-alpha-b 828-836 (SEQ ID NO: 2249) A*02: 01 YLIELIDRV TACE 250-258 (SEQ ID NO: 2250) A*02: 01 FLAEDALNTV Epithelial Discoidin Domain Receptor 1 (SEQ ID NO: 2251) (EDDR1) 867-876 A*02: 01 GLMKYIGEV TRPM8 187-195 (SEQ ID NO: 2252) A*02: 01 AILALLPAL Prostate Stem Cell Antigen (PSCA) (SEQ ID NO: 2253) 105-133 A*02: 01 GLQHWVPEL BA46 (Lactadherin) 97-106 (SEQ ID NO: 2254) A*02: 01 GVRGRVEEI BCR-ABL (SEQ ID NO: 2255) A*02: 01 ITDQVPFSV gp100 (pmel) 209-217 (SEQ ID NO: 2256) A*02: 01 KLCPVQLWV p53 139-147 (SEQ ID NO: 2257) A*02: 01 KVAEELVHFL MAGEA3 112-120 (alternative version) (SEQ ID NO: 2258) A*02: 01 SLPPPGTRV p53 149-157 (SEQ ID NO: 2259) A*02: 01 YLGSYGFRL p53 103-111 (SEQ ID NO: 2260) A*02: 01 YLQLVFGIEV MAGEA2 157-166 (SEQ ID NO: 2261) A*02: 01 TLQDIVYKL BMI1 74-82 (SEQ ID NO: 2262) A*02: 01 YAIDLPVSV L-dopachrome tautomerase 488-496 (SEQ ID NO: 2263) A*02: 01 AMVGAVLTA Tyrosinase 482-190 (SEQ ID NO: 2264) A*02: 01 ATVGIMIGV CEACAM5 687-695 (SEQ ID NO: 2265) A*02: 01 YVDPVITSI Hepatocyte growth factor receptor (SEQ ID NO: 2266) 673-681 A*02: 01 GVLLWEIFSL VEGFR1 28-37 (SEQ ID NO: 2267) A*02: 01 LMAQEALAFL CAMEL 2-11 (SEQ ID NO: 2268) A*02: 01 RVA(PHOSPHO-S)PTSGV Insulin receptor substrate-2 1097-1105 (SEQ ID NO: 2269) A*02: 01 RVASPTSGV IRS-2 1097-1105 (SEQ ID NO: 2270) A*02: 01 ALNVYNGLL ACPP 299-307 (SEQ ID NO: 2271) A*02: 01 ALSPVPPVV Bcl-2 85-93 (SEQ ID NO: 2272) A*02: 01 ALVCYGPGI FAP alpha 463-471 (SEQ ID NO: 2273) A*02: 01 ALWPWLLMAT RNF43 11-20 (SEQ ID NO: 2274) A*02: 01 ALYLMELTM CB9L2 (SEQ ID NO: 2275) A*02: 01 CLPSPSTPV BMI1 271-279 (SEQ ID NO: 2276) A*02: 01 ELSDSLGPV PASD1 695-703 (SEQ ID NO: 2277) A*02: 01 FLFLRNFSL TARP(V28L)27-35 (SEQ ID NO: 2278) A*02: 01 FLPSPLFFFL TARP(PSL)5-13 (SEQ ID NO: 2279) A*02: 01 GLFKCGIAV FAP 639-647 (SEQ ID NO: 2280) A*02: 01 GLIQLVEGV TRAG-3 4-12 (SEQ ID NO: 2281) A*02: 01 ILGVLTSLV DLK1 309-317 (SEQ ID NO: 2282) A*02: 01 LLVPTCVFLV 691-700 (SEQ ID NO: 2283) A*02: 01 MLAVFLPIV STEAP 292-300 (293L) (SEQ ID NO: 2284) A*02: 01 NLFETPVEA 194-202 (SEQ ID NO: 2285) A*02: 01 QLGEQCWTV PSCA 44-51 (51A) (SEQ ID NO: 2286) A*02: 01 RLAEYQAYI SART3 309-317 (SEQ ID NO: 2287) A*02: 01 SIDWFMVTV p31-39 (SEQ ID NO: 2288) A*02: 01 SILLRDAGLV TRAG-3 57-66 (SEQ ID NO: 2289) A*02: 01 SLFEPPPPG PSMA 85-93 (SEQ ID NO: 2290) A*02: 01 SQADALKYV EZH2 729-737 (SEQ ID NO: 2291) A*02: 01 WLSLKTLLSL Bcl-2 214-223 (SEQ ID NO: 2292) A*02: 01 YLNRHLHTWI BCL-2 180-189 (SEQ ID NO: 2293) A*02: 01 YLQWIEFSI Prominin1 744-752 (SEQ ID NO: 2294) A*02: 01 YLYQWLGAPV Osteocalcin 51-60 (SEQ ID NO: 2295) A*02: 01 KLMSSNSTDL HSP105 234-243 (SEQ ID NO: 2296) A*02: 01 RLQGISPKI SSX2 103-111 (SEQ ID NO: 2297) A*02: 01 AILALLPALL PSCA (SEQ ID NO: 2298) A*02: 01 ALIHHNTHL HER2 466-474 (SEQ ID NO: 2299) A*02: 01 CMHLLLEAV MG50 624-632 (SEQ ID NO: 2300) A*02: 01 FLIIWQNTM F5P26 (SEQ ID NO: 2301) A*02: 01 FLPWHRLFLL Tyrosinase 207-216 (SEQ ID NO: 2302) A*02: 01 FVWLHYYSV TRP2 185-193(L) (SEQ ID NO: 2303) A*02: 01 GLFGDIYLA CSNK1A1 26-34 (SEQ ID NO: 2304) A*02: 01 GLFGDIYLAI CSNK1A1 26-35 (SEQ ID NO: 2305) A*02: 01 ILLRDAGLV TRAG-3L 58-66 (SEQ ID NO: 2306) A*02: 01 ILLVVVLGV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2307) 707-715 A*02: 01 ILNAMIAKI HAUS3 154-162 (SEQ ID NO: 2308) A*02: 01 KASEYLQLV MAGEA2 153-161 (SEQ ID NO: 2309) A*02: 01 KIWEELSVL MAGEA3 220-228 (SEQ ID NO: 2310) A*02: 01 KLIDRTE(S)L LSP1 325-333 (SEQ ID NO: 2311) A*02: 01 KLTGDENFTI Tyrosinase precursor 224-233 (SEQ ID NO: 2312) A*02: 01 LLCYSCKAQV PSCA 17-26 (SEQ ID NO: 2313) A*02: 01 LLLEAVPAV MG50 69-77 (SEQ ID NO: 2314) A*02: 01 LLNQLQVNL Mucin2 467-475 (SEQ ID NO: 2315) A*02: 01 LLRDAGLVKM TRAP 59-68 (SEQ ID NO: 2316) A*02: 01 LLRRYNVAKV SOX11 266-275 (SEQ ID NO: 2317) A*02: 01 LLSHGAVIEV Ankyrin NYBR1 158-167 (SEQ ID NO: 2318) A*02: 01 LVFGIELMEV MAGEA3 160-169 (SEQ ID NO: 2319) A*02: 01 LVFGIEVVEV MAGEA12 160-169 (SEQ ID NO: 2320) A*02: 01 MLWGWREHV Mucin2 645-653 (SEQ ID NO: 2321) A*02: 01 PLQPEQLQV Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2322) 437-445 A*02: 01 QLMAFNHLI PAX3/FKHR 135-143 (SEQ ID NO: 2323) A*02: 01 QLMPYGCLL Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2324) 845-853 A*02: 01 RLGPTLMCL MG50 1244-1252 (SEQ ID NO: 2325) A*02: 01 RLTRFLSRV CyclinD 228-236 (SEQ ID NO: 2326) A*02: 01 RTF(S)PTYGL Desmuslin 426-434 (SEQ ID NO: 2327) A*02: 01 SILLRDAGL TRAP 57-65 (SEQ ID NO: 2328) A*02: 01 SLADEAEVYL GAS7 Neoepitope (SEQ ID NO: 2329) A*02: 01 SLDDYNHLV L-dopachrome tautomerase 288-296 (SEQ ID NO: 2330) A*02: 01 SLYKFSPFPL O-linked N-acetylglucosamine transferase (SEQ ID NO: 2331) FSP06 A*02: 01 SMTR(S)PPRV SFRS2B 241-249 (SEQ ID NO: 2332) A*02: 01 TLEEITGYL Receptor tyrosine-protein kinase erbB-2 (SEQ ID NO: 2333) 448-456 A*02: 01 TLHCDCEIL MG50 210-218 (SEQ ID NO: 2334) A*02: 01 VLEPPGARDV BIR 7 230-239 (SEQ ID NO: 2335) A*02: 01 VLLALLMAGL Prostate stem cell antigen 4-13 (SEQ ID NO: 2336) A*02: 01 VLSVNVPDV MG50 625-633 (SEQ ID NO: 2337) A*02: 01 VLVKSPNHV Receptor tyrosine-protein kinase erbB-4 890-898 (SEQ ID NO: 2338) A*02: 01 VMIG(S)PKKV Tensin3 1558-1566 (SEQ ID NO: 2339) A*02: 01 VVLGVVFGI Receptor tyrosine-protein kinase erbB-2 743-751 (SEQ ID NO: 2340) A*02: 01 WLPKILGEV MG50 1051-1059 (SEQ ID NO: 2341) A*02: 01 WLQYFPNPV Cytochrome P450 246-254 (SEQ ID NO: 2342) A*02: 01 YLLDLSTNHL Fibromodulin 7-15 (SEQ ID NO: 2343) A*02: 01 YLWWVNNQSL CEA 176-185 (SEQ ID NO: 2344) A*02: 01 ALGGHPLLGV Dickkopf-related protein 1 20-29 (SEQ ID NO: 2345) A*02: 01 ALLAGLVSL FGFR4 676-684 (SEQ ID NO: 2346) A*02: 01 ALLTYMIAHI Thymidylate synthase 231-240 (SEQ ID NO: 2347) A*02: 01 ALMDKSLHV MART-1 56-64 (SEQ ID NO: 2348) A*02: 01 ALPPPLMLL Heparanase 8-16 (SEQ ID NO: 2349) A*02: 01 ALSVMGVYV MAGEA9 223-231 (SEQ ID NO: 2350) A*02: 01 ALVEFEDVL hnRNP L 140-148 (SEQ ID NO: 2351) A*02: 01 ALWPWLLMA RNF43 11-19 (SEQ ID NO: 2352) A*02: 01 AMLGTHTMEV Melanocyte-specific secreted glycoprotein 184-193 (SEQ ID NO: 2353) A*02: 01 AVIGALLAV Melanocyte-specific secreted glycoprotein 20-28 (SEQ ID NO: 2354) A*02: 01 CLYGNVEKV hnRNP L 404-412 (SEQ ID NO: 2355) A*02: 01 DLIFGLNAL Heparanase 185-193 (SEQ ID NO: 2356) A*02: 01 ELFQDLSQL ETV5 54-53 (SEQ ID NO: 2357) A*02: 01 FAWERVRGL Cyclin-dependent kinase inhibitor 1 97-105 (SEQ ID NO: 2358) A*02: 01 FIASNGVKLV ACTN4 118-127 (K5N) (SEQ ID NO: 2359) A*02: 01 FLALIICNA Tubulin beta 4 283-291 (SEQ ID NO: 2360) A*02: 01 FLDEFMEGV Malic enzyme 224-232 (SEQ ID NO: 2361) A*02: 01 RMFPNAPYL WT-1 126-134 (Wilms tumor) (SEQ ID NO: 2362) A*02: 01 RLNMFTPYI Chlamydia trachomatis MOMP 258-266 (SEQ ID NO: 2363) A*02: 01 NMFTPYIGV MOMP precursor 283-291 (SEQ ID NO: 2364) A*02: 01 NLVPMVATV HCMV pp65 495-504 (SEQ ID NO: 2365) A*02: 01 VLEETSVML HCMVIE1 316-324 (UL123) (SEQ ID NO: 2366) A*02: 01 VLAELVKQI HCMVIE1 81-89 (SEQ ID NO: 2367) A*02: 01 MLNIPSINV pp65 120-128 (SEQ ID NO: 2368) A*02: 01 LLLDRLNQL SARS-CoV Nucleocapsid protein 223-231 (SEQ ID NO: 2369) (conserved in SARS-CoV-2) A*02: 01 FIAGLIAIV SARS-CoV-2 Spike glycoprotein 1220-1228 (SEQ ID NO: 2370) (confirmed epitope) A*02: 01 ALNTLVKQL SARS-CoV Spike glycoprotein precursor (SEQ ID NO: 2371) 940-948 (conserved in SARS-CoV-2) A*02: 01 LITGRLQSL SARS-CoV-2 Spike glycoprotein 996-1004 (SEQ ID NO: 2372) (confirmed epitope) A*02: 01 NLNESLIDL SARS-CoV (SEQ ID NO: 2373) Spike glycoprotein precursor 1174-1182 (conserved in SARS-Cov-2) A*02: 01 VLNDILSRL SARS-CoV Spike glycoprotein precursor (SEQ ID NO: 2374) 958-966 (conserved in SARS-Cov-2) A*02: 01 YLQPRTFLL SARS-Cov-2 Spike glycoprotein 269-277 (SEQ ID NO: 2375) (confirmed epitope) A*02: 01 LLYDANYFL SARS-CoV-2 ORF3a 139-147 (confirmed epitope) (SEQ ID NO: 2376) A*02: 01 RLQSLQTYV SARS-CoV-2 Spike glycoprotein 1000-1008 (SEQ ID NO: 2377) (confirmed subdominant epitope) A*02: 01 KLWAQCVQL SARS-CoV-2 ORF1ab 3886-3894 (confirmed epitope) (SEQ ID NO: 2378) A*02: 01 TLYAVATTI Dengue NS4b 40-48 (SEQ ID NO: 2379) A*02: 01 KLAEAIFKL Dengue NS5 563-571 (SEQ ID NO: 2380) A*02: 01 ILIRTGLLVI Dengue NS2b 97-106 (SEQ ID NO: 2381) A*02: 01 AIKRGLRTL Dengue NS3 112-120 (SEQ ID NO: 2382) A*02: 01 LLLGLMILL Dengue NS4a 56-64 (SEQ ID NO: 2383) A*02: 01 VLLLVTHYA Dengue NS4b 111-119 (SEQ ID NO: 2384) A*02: 01 GLCTLVAML EBV BM LF-1 259-267 (SEQ ID NO: 2385) A*02: 01 CLGGLLTMV EBV LMP-2 426-434 (SEQ ID NO: 2386) A*02: 01 YLLEMLWRL EBV LMP-1 125-133 (SEQ ID NO: 2387) A*02: 01 YLQQNWWTL EBV LMP1 159-167 (SEQ ID NO: 2388) A*02: 01 YVLDHLIVV EBV BRLF1 109-117 (SEQ ID NO: 2389) A*02: 01 FLYALALLL EBV LMP-2 356-364 (SEQ ID NO: 2390) A*02: 01 TLDYKPLSV EBV BMRF1 208-216 (SEQ ID NO: 2391) A*02: 01 LLDFVRFMGV EBV EBNA-3C 284-293 (SEQ ID NO: 2392) A*02: 01 FLDKGTYTL EBV BALF-4 276-284 (SEQ ID NO: 2393) A*02: 01 FLPSDFFPSV HBV core antigen 18-27 (SEQ ID NO: 2394) A*02: 01 FLLTRILTI HBV envelope 183-191 (SEQ ID NO: 2395) A*02: 01 GLSPTVWLSV HBV surface antigen 185-194 (SEQ ID NO: 2396) A*02: 01 WLSLLVPFV HBV surface antigen 172-181 (SEQ ID NO: 2397) A*02: 01 FLLSLGIHL HBV polymerase 573-581 (SEQ ID NO: 2398) A*02: 01 FLPSDFFPSI HBV core 18-27 (subtype ADR4) (SEQ ID NO: 2399) A*02: 01 VLHKRTLGL HBV X 92-100 (SEQ ID NO: 2400) A*02: 01 GLSRYVARL HBV Pol 455-463 (SEQ ID NO: 2401) A*02: 01 YMDDVVLGA HBV Polymerase 548-556 (SEQ ID NO: 2402) A*02: 01 KLHLYSHPI HBV Pol 502-510 (SEQ ID NO: 2403) A*02: 01 ELMTLATWV HBV core protein 64-72 (SEQ ID NO: 2404) A*02: 01 DLMGYIPAV HCV core 132-140 (SEQ ID NO: 2405) A*02: 01 CINGVCWTV HCV NS3 1073-1081 (SEQ ID NO: 2406) A*02: 01 YLLPRRGPRL HCV core 35-44 (SEQ ID NO: 2407) A*02: 01 VLSDFKTWL HCV NS5a 1987-1995 (SEQ ID NO: 2408) A*02: 01 ALYDVVTKL HCV NS5b 2594-2602 (SEQ ID NO: 2409) A*02: 01 KLVALGINAV HCV NS3 1406-1415 (SEQ ID NO: 2410) A*02: 01 LLFNILGGWV HCV NS4b 1807-1816 (SEQ ID NO: 2411) A*02: 01 KLSGLGINAV HCV NS3 1406-1415 (SEQ ID NO: 2412) A*02: 01 DLMGYIPLV HCV core 132-140 (SEQ ID NO: 2413) A*02: 01 CVNGVCWTV HCV NS3 1073-1081 (SEQ ID NO: 2414) A*02: 01 GLQDCTMLV HCV NS5B 2727-2735 (SEQ ID NO: 2415) A*02: 01 SLYNTVATL HIV-1 gag p17 76-84 (SEQ ID NO: 416) A*02: 01 ILKEPVHGV HIV-1 RT476-484 (SEQ ID NO: 2417) A*02: 01 TLNAWVKVV HIV-1 gag p24 19-27 (SEQ ID NO: 2418) A*02: 01 KLTPLCVTL HIV-1 env gp120 90-98 (SEQ ID NO: 2419) A*02: 01 GLADQUHL HIV-1 vif 101-109 (SEQ ID NO: 2420) A*02: 01 LTFGWCFKL HIV-1 nef 137-145 (SEQ ID NO: 2421) A*02: 01 FLGKIWPS Gag 433-440 (SEQ ID NO: 2422) A*02: 01 ALVEMGHHA HIV Vpu 66-74 (SEQ ID NO: 2423) A*02: 01 RTLNAWVKV HIV gag 150-158 (SEQ ID NO: 2424) A*02: 01 NVWATHACV HIV env gp 67-7 (SEQ ID NO: 2425) A*02: 01 SLLNATAIAV HIV env 816-825 (SEQ ID NO: 2426) A*02: 01 SLFNTVATL HIV gag 77-85 (SEQ ID NO: 2427) A*02: 01 SLVKHHMYI HIV vif 23-31 (SEQ ID NO: 2428) A*02: 01 VIYHYVDDL HIV pol (SEQ ID NO: 2429) A*02: 01 YMLDLQPETT HPV 16 E7 11-20 (SEQ ID NO: 2430) A*02: 01 KLPQLCTEL HPV 16 E6 18-26 (SEQ ID NO: 2431) A*02: 01 YMLDLQPET HPV 16 E7 11-19 (SEQ ID NO: 2432) A*02: 01 MLDLQPETT HPV 16 E7 12-20 (SEQ ID NO: 2433) A*02: 01 VLMIKALEL Non muscle Myosin-9 741-749 (SEQ ID NO: 2434) A*02: 01 QLFNHTMFI Non-muscle Myosin 478-486 (SEQ ID NO: 2435) A*02: 01 QMARLAWEA 1116-1124 (SEQ ID NO: 2436) A*02: 01 LLFGYPVYV Human T-cell lymphotropic virus-1 (HTLV-1) tax 11-19 (SEQ ID NO: 2437) A*02: 01 AVLDGLLSL HTLV bZIP factor 42-50 (SEQ ID NO: 2438) A*02: 01 GLLSLEEEL bZIP factor 26-34 (SEQ ID NO: 2439) A*02: 01 GILGFVFTL Influenza A MP 58-66 (SEQ ID NO: 2440) A*02: 01 ILGFVFTLTV Influenza A MP 59-68 (SEQ ID NO: 2441) A*02: 01 KLGEFYNQMM Flu BNP 85-94 (Influenza B) (SEQ ID NO: 2442) A*02: 01 SITEVECFL VP1 36-44 (SEQ ID NO: 2443) A*02: 01 ILMWEAVTL VP1 100-108 (SEQ ID NO: 2444) A*02: 01 ALPHIIDEV LCMV envelope gp 10-18 (SEQ ID NO: 2445) A*02: 01 YLVSIFLHL LCMV envelope gp 447-455 (SEQ ID NO: 2446) A*02: 01 SLNQTVHSL NP 69-77 (SEQ ID NO: 2447) A*02: 01 YLNKIQNSL Plasmodium falciparum CSP 334-342 (SEQ ID NO: 2448) A*02: 01 FIDSYICQV miHAg H-Y (human SMCY) 311-319 (SEQ ID NO: 2449) A*02: 01 YIGEVLVSV HA-2 (SEQ ID NO: 2450) A*02: 01 VLHDDLLEA Minor Histocompatibility Antigen HA-1 137-145 (SEQ ID NO: 2451) A*02: 01 RTLDKVLEV miHAg HA-8 (SEQ ID NO: 2452) A*02: 01 NEGATIVE Negative Control (SEQ ID NO: 2453) A*02: 01 TMFPHIIVDV Norovirus VP1 139-148 (SEQ ID NO: 2454) A*02: 01 LLDVPTAAV Interferon gamma inducible protein (GILT) 30 27-35 (SEQ ID NO: 2455) A*02: 01 RILGAVAKV Vinculin 822-830 (SEQ ID NO: 2456) A*02: 01 LMWYELSKI KSHVF-8 g6.492-500 (SEQ ID NO: 2457) A*02: 01 ILEDIVLTL Streptococcus pyogenes Cas9 615-623 (SEQ ID NO: 2458) A*02: 01 KMLKEMGEV RSV NP 137-145 (SEQ ID NO: 2459) A*02: 01 KLIANNTRV Mycobacterium bovis antigen 85-A 200-208 (SEQ ID NO: 2460) A*02: 01 GLPVEYLQV Mycobacterium bovis antigen 85-A 6-14 (SEQ ID NO: 2461) A*02: 01 GILTVSVAV 16 kDa (SEQ ID NO: 2462) A*02: 01 AMASTEGNV ESAT-6 (SEQ ID NO: 2463) A*02: 01 VLTDGNPPEV 19 kDa (SEQ ID NO: 2464) A*02: 01 KVDDTFYYV Vaccinia virus Host range protein 2 74-82 (SEQ ID NO: 2465) A*02: 01 ILDDNLYKV Vaccinia virus Copenhagen Protein G5 18-26 (SEQ ID NO: 2466) A*02: 01 ALWALPHAA 1E62 593-601 (SEQ ID NO: 2467) A*02: 01 RLDDDGNFQL West Nile Virus NY-99 polyprotein precursor (SEQ ID NO: 2468) (1452-1461) A*02: 01 ATWAENIQV West Nile virus NY-99 polyprotein precursor 3390-3398 (SEQ ID NO: 2469) A*02: 01 YTMDGEYRL West Nile virus NY-99 polyprotein precursor 2023-2031 (SEQ ID NO: 2470) A*02: 01 SVGGVFTSV WNV envelope gp 430-438 (SEQ ID NO: 2471) A*02: 01 SLFGQRIEV WNV nonstructural protein 4B 15-23 (SEQ ID NO: 2472) A*02: 01 LLWNGPMAV NS4B 214-222 (SEQ ID NO: 2473) A*03: 01 KQSSKALQR bcr-abl 210 kD fusion protein 21-29 (SEQ ID NO: 2474) A*03: 01 ALLAVGATK gp100 (pmel17) 17-25 (SEQ ID NO: 2475) A*03: 01 ATGFKQSSK bcr-abl 210 kD fusion protein 259-269 (SEQ ID NO: 2476) A*03: 01 RISTFKNWPK Survivin-3A 18-27 (27K) (SEQ ID NO: 2477) A*03: 01 RLGLQVRKNK RhoC 176-185 (1770 (SEQ ID NO: 2478) A*03: 01 RLLFFAPTR Mcl-1 95-103 (SEQ ID NO: 2479) A*03: 01 QVLKKIAQK HMOX1 145-153 (SEQ ID NO: 2480) A*03: 01 RIAAWMATY 165-173 (SEQ ID NO: 2481) A*03: 01 KLGGALQAK HCMVIE1 184-192 (SEQ ID NO: 2482) A*03: 01 KTFPPTEPK SARS-CoV-2 Nucleocapsid protein 362-370 (SEQ ID NO: 2483) (confirmed epitope) A*03: 01 ELERAADVK Dengue NS2b 52-60 (SEQ ID NO: 2484) A*03: 01 RVSTVQQLTK Dengue C 22-31 (SEQ ID NO: 2485) A*03: 01 RIEPSWADVK Dengue NS3 64-74 (SEQ ID NO: 2486) A*03: 01 RVIDPRRCMK Dengue NS3 422-431 (SEQ ID NO: 2487) A*03: 01 KITAEWLWK Dengue NS5 375-383 (SEQ ID NO: 2488) A*03: 01 RLRAEAQVK EBV EBNA 3A 603-611 (SEQ ID NO: 2489) A*03: 01 RVRAYTYSK EBV BRLF1 (SEQ ID NO: 2490) A*03: 01 RVCEKMALY HCV NS5B 2588-2596 (SEQ ID NO: 2491) A*03: 01 QVPLRPMTYK HIV-1 nef 73-82 (SEQ ID NO: 2492) A*03: 01 RLRPGGKKK HIV-1 gag p17 19-27 (SEQ ID NO: 2493) A*03: 01 AIFQSSMTK HIV pol 325-333 (SEQ ID NO: 2494) A*03: 01 KLCLRFLSK HPV 33 E6 64-72 (SEQ ID NO: 2495) A*03: 01 ILRGSVAHK Influenza A (PR8) NP 265-274 (SEQ ID NO: 2496) A*11: 01 KTFPPTEPK SARS-CoV-2 Nucleocapsid protein 362-370 (SEQ ID NO: 2483) (confirmed epitope) A*11: 01 GTSGSPIINR Dengue NS3 serotype 3&4 133-142 (SEQ ID NO: 2497) A*11: 01 GTSGSPIIDK Dengue NS3 133-142 (SEQ ID NO: 2498) A*11: 01 GTSGSPIVNR NS3 serotype 1 133-142 (SEQ ID NO: 2499) A*11: 01 GTSGSPIVDR Dengue NS3 serotype 2 133-142 (SEQ ID NO: 2500) A*11: 01 GTSGSPIADK Dengue NS3 133-142 (SEQ ID NO: 2501) A*11: 01 RVSTVQQLTK Dengue C 22-31 (SEQ ID NO: 2485) A*11: 01 RIEPSWADVK Dengue NS3 64-74 (SEQ ID NO: 2486) A*11: 01 RVIDPRRCMK Dengue NS3 422-431 (SEQ ID NO: 2487) A*11: 01 KITAEWLWK Dengue NS5 375-383 (SEQ ID NO: 2488) A*11: 01 IVTDFSVIK EBV EBNA-4 416-424 (SEQ ID NO: 2502) A*11: 01 SSCSSCPLSK EBV LMP-2 340-349 (SEQ ID NO: 2503) A*11: 01 ATIGTAMYK EBV BRLF1 134-142 (SEQ ID NO: 2504) A*11: 01 AVFDRKSDAK EBNA3B 399-408 (SEQ ID NO: 2505) A*11: 01 YVNVNMGLK HBV core antigen 88-96 (SEQ ID NO: 2506) A*11: 01 YVNTNMGLK HBV core 88-96 (SEQ ID NO: 2507) A*11: 01 STLPETTVVRR HBV core 141-151 (SEQ ID NO: 2508) A*11: 01 AVDLSHFLK HIV nef 84-92 (SEQ ID NO: 2509) A*11: 01 ACQGVGGPGHK HIV gag p24 (SEQ ID NO: 2510) A*11: 01 NTLEQTVKK HPV 33 E6 86-94 (SEQ ID NO: 2511) A*11: 01 SIIPSGPLK Influenza A MP 13-21 (SEQ ID NO: 2512) A*11: 01 RMVLASTTAK Influenza A MP1 178-187 (SEQ ID NO: 2513) A*11: 01 KSMREEYRK Influenza A MP2 70-78 (SEQ ID NO: 2514) A*24: 02 TYFSLNNKF Adenovirus 5 Hexon 37-45 (SEQ ID NO: 2515) A*24: 02 TYACFVSNL Carcinogenic Embryonic Antigen (CEA) 652-660 (SEQ ID NO: 2516) A*24: 02 AFLPWHRLF Tyrosinase 188-196 (SEQ ID NO: 2517) A*24: 02 IMPKAGLLI MAGE-A3 (SEQ ID NO: 2518) A*24: 02 VYFFLPDHL gp100-intron 4 (170-178) (SEQ ID NO: 2519) A*24: 02 EYLQLVFGI MAGEA2 156-164 (SEQ ID NO: 2520) A*24: 02 TYLPTNASL HER-2/neu 63-71 (SEQ ID NO: 2521) A*24: 02 VYGFVRACL Telomerase reverse transcriptase (hTRT) 461-469 (SEQ ID NO: 2522) A*24: 02 TFPDLESEF MAGEA3 97-105 (SEQ ID NO: 2523) A*24: 02 DYLQYVLQI MiHA ACC1 15-23 (SEQ ID NO: 2524) A*24: 02 RYCNLEGPPI Lymphocyte antigen 6 complex locus K (LY6K) 177-186 (SEQ ID NO: 2525) A*24: 02 AYACNTSTL Survivin 80-88 (SEQ ID NO: 2526) A*24: 02 CYASGWGSI Prostate Specific Antigen-1 153-161 (SEQ ID NO: 2527) A*24: 02 DYLNEWGSRF CDH3 807-816 (SEQ ID NO: 2528) A*24: 02 EYCPGGNLF MELK 87-95 (93N) (SEQ ID NO: 2529) A*24: 02 EYYELFVNI DEP DC1 294-302 (SEQ ID NO: 2530) A*24: 02 GYCTQIGIF HENMT1 221-229 (SEQ ID NO: 2531) A*24: 02 IYTWIEDHF FOXM1 262-270 (SEQ ID NO: 2532) A*24: 02 NYQPVWLCL RNF43 721-729 (722Y) (SEQ ID NO: 2533) A*24: 02 RYNAQCQETI Midkine 110-119 (SEQ ID NO: 2534) A*24: 02 EYRALQLHL CA9 219-227 (SEQ ID NO: 2535) A*24: 02 SYRNEIAYL TTK protein kinase 551-559 (SEQ ID NO: 2536) A*24: 02 VYLRVRPLL KIF20A 67-75 (SEQ ID NO: 2537) A*24: 02 VYYNWQYLL IL13r 146-154 (SEQ ID NO: 2538) A*24: 02 VYALPLKML HCMV pp65 113-121 (SEQ ID NO: 2539) A*24: 02 QYDPVAALF HCMV pp65 341-349 (SEQ ID NO: 2540) A*24: 02 AYAQKIFKI CMV 1E-1 248-256 (SEQ ID NO: 2541) A*24: 02 QYSDRRWCF Dengue NS3 557-565 (Singapore/S275/1990) (SEQ ID NO: 2542) A*24: 02 TYGPVFMCL EBV LMP-2 419-427 (SEQ ID NO: 2543) A*24: 02 PYLFWLAAI EBV LMP2 131-139 (SEQ ID NO: 2544) A*24: 02 TYGPVFMSL EBV LMP2 419-427 (SEQ ID NO: 2545) A*24: 02 EYLVSFGVW HBV core 117-125 (SEQ ID NO: 2546) A*24: 02 KYTSFPWLL HBV polymerase 756-764 (SEQ ID NO: 2547) A*24: 02 FFPSIRDLL HBV core protein 23-31 (SEQ ID NO: 2548) A*24: 02 AYSQQTRGL HCV NS3 1031-1039 (SEQ ID NO: 2549) A*24: 02 RYPLTFGWCY HIV-1 Nef 134-143 (SEQ ID NO: 2550) A*24: 02 RYLKDQQLL HIV-1 gag gp41 67-75 (SEQ ID NO: 2551) A*24: 02 RYLRDQQLL HIV env (SEQ ID NO: 2552) A*24: 02 RYPLTFGWCF HIV nef 143-152 (SEQ ID NO: 2553) A*24: 02 RYPLTFGW HIV nef (SEQ ID NO: 2554) A*24: 02 VYDFAFRDL HPV16 E6 (SEQ ID NO: 2555) A*24: 02 SFHSLHLLF HTLV Tax 301-309 (SEQ ID NO: 2556) A*29: 02 KEKYIDQEEL HSP90 alpha 280-288 (Pathologic Conditions) (SEQ ID NO: 2557) A*29: 02 LYNTVATLY HIV gag 79-86 (SEQ ID NO: 2558) A*29: 02 SFDPIPIHY HIV env 216-224 (SEQ ID NO: 2559) A*29: 02 SFNCRGEFFY HIV env 382-391 (SEQ ID NO: 2560) A*68: 01 TVSGNILTIR NY-ESO-1 127-136 (SEQ ID NO: 2561) B*07: 02 VPQYGYLTL AAV2 372-380 (SEQ ID NO: 2562) B*07: 02 KPYSGTAYNSL Adenovirus Hexon 114-124 (SEQ ID NO: 2563) B*07: 02 KPYSGTAYNAL Adenovirus Hexon 114-124 (SEQ ID NO: 2564) B*07: 02 LPLMRKAYL LT antigen 27-35 (SEQ ID NO: 2565) B*07: 02 LPWHRLFLL Tyrosinase 208-216 (SEQ ID NO: 2566) B*07: 02 EPR(PHOSPHO-S)PSHSM Insulin receptor substrate 2 (SEQ ID NO: 2567) B*07: 02 TPNQRQNVC P2X5 (SEQ ID NO: 2568) B*07: 02 APRGVRMAV LAGE-1 46-54 (SEQ ID NO: 2569) B*07: 02 LPVSPRLQL CEACAM 185-193 (SEQ ID NO: 2570) B*07: 02 TPRVTGGGAM HCMV pp65 417-426 (SEQ ID NO: 2571) B*07: 02 RPHERNGFTVL HCMV pp65 265-275 (SEQ ID NO: 2572) B*07: 02 SPRWYFYYL SARS-CoV-2 Nucleocapsid protein 105-113 (SEQ ID NO: 2573) (confirmed epitope) B*07: 02 APTRVVAAEM Dengue NS3 serotype 2 222-231 (SEQ ID NO: 2574) B*07: 02 RPPIFIRRL EBV EBNA-3A 247-255 (SEQ ID NO: 2575) B*07: 02 RPQGGSRPEFVKL EBV BMRF1 116-128 (SEQ ID NO: 2576) B*07: 02 QPRAPIRPI EBV EBNA-3C 881-889 (SEQ ID NO: 2577) B*07: 02 LPSDFFPSV HBV core 19-27 (SEQ ID NO: 2578) B*07: 02 GPRLGVRAT HCV core 41-49 (SEQ ID NO: 2579) B*07: 02 DPRRRSRNL HCV core 111-119 (SEQ ID NO: 2580) B*07: 02 IPRRIRQGL HIV-1 env gp120 848-856 (SEQ ID NO: 2581) B*07: 02 TPGPGVRYPL HIV-1 nef 128-137 (SEQ ID NO: 2582) B*07: 02 GPGHKARVL HIV gag p24 223-231 (SEQ ID NO: 2583) B*07: 02 KPTLKEYVL HPV 33 E7 5-13 (SEQ ID NO: 2584) B*07: 02 LPVSCPEDL bZIP factor 10-18 (SEQ ID NO: 2585) B*07: 02 QPEWFRNVL Influenza A PB1 329-337 (SEQ ID NO: 2586) B*07: 02 SPIVPSFDM Influenza A NP 473-481 (SEQ ID NO: 2587) B*07: 02 SPSVDKARAEL MiHAg SMCY 1041-1051 (SEQ ID NO: 2588) B*08: 01 GFKQSSKAL bcr-abl 210 kD fusion protein 19-27 (SEQ ID NO: 2589) B*08: 01 ELRRKMMYM 1E1 199-207 (SEQ ID NO: 2590) B*08: 01 ELKRKMIYM CMV 1E-1 199-207 (RA Position 3, M/I Position 7) (SEQ ID NO: 2591) B*08: 01 RIKQKGIL Dengue NS3 25-32 (SEQ ID NO: 2592) B*08: 01 LEKTKKDL Dengue NS4a 6-13 (SEQ ID NO: 2593) B*08: 01 FLRGRAYGL EBV EBNA-3A 193-201 (SEQ ID NO: 2594) B*08: 01 RAKFKQLL EBV BZLF-1 190-197 (SEQ ID NO: 2595) B*08: 01 QAKWRLQTL EBV EBNA3A 158-166 (SEQ ID NO: 2596) B*08: 01 GLKILQLL HBV external core Ag (SEQ ID NO: 2597) B*08: 01 HSKKKCDEL HCV NS3 1395-1403 (SEQ ID NO: 2598) B*08: 01 FLKEKGGL HIV-1 nef 90-97 (SEQ ID NO: 2599) B*08: 01 GEIYKRWII HIV-1 gag p24 261-269 (SEQ ID NO: 2600) B*08: 01 ElYKRWII HIV p24 gag 128-135 (SEQ ID NO: 2601) B*08: 01 YLKDQQLL Env 586-593 (SEQ ID NO: 2602) B*15: 01 CLIPTAMAF Dengue C 107-115 (SEQ ID NO: 2603) B*15: 01 RLRPGGKKKY HIV-1 p17 20-29 (SEQ ID NO: 2604) B*27: 05 GRFGLATEK BRAF 594-601 (600E) (SEQ ID NO: 2605) B*27: 05 GRFGLATVK BRAF594-601 (600V) (SEQ ID NO: 2606) B*27: 05 RMFPNAPYL WT-1 126-134 (Wilms tumor) (SEQ ID NO: 2362) B*27: 05 ARKLLLDNL PqqC-like protein 70-78 (SEQ ID NO: 2607) B*27: 05 NRAKQVIKL Probable ATP-dependent Clp protease (SEQ ID NO: 2608) ATP-binding subunit 7-15 B*27: 05 QRNAPRITF SARS-CoV-2 Nucleocapsid protein 9-17 (SEQ ID NO: 2609) (confirmed epitope) B*27: 05 ARMILMTHF HCV NS5B 2841-2849 (SEQ ID NO: 2610) B*27: 05 KRWIILGLNK HIV-1 gag p24 265-274 (SEQ ID NO: 2611) B*27: 05 KRWIIMGLNK HIV-1 Gag p24 263-272 (SEQ ID NO: 2612) B*27: 05 GRAFVTIGK HIV-1 gp100 103-111 (SEQ ID NO: 2613) B*27: 05 SRYWAIRTR Influenza A NP 383-391 (SEQ ID NO: 2614) B*35: 01 IPYLDGTFY Adenovirus Hexon (SEQ ID NO: 2615) B*35: 01 MPFATPMEA NY-ESO-1 94-102 (SEQ ID NO: 2616) B*35: 01 IPSINVHHY HCMV pp65 123-131 (SEQ ID NO: 2617) B*35: 01 TPEGIIPTL Dengue NS3 500-508 (SEQ ID NO: 2618) B*35: 01 YPLHEQHGM EBV EBNA-3A 458-466 (SEQ ID NO: 2619) B*35: 01 EPLPQGQLTAY EBV BZLF-1 54-64 (SEQ ID NO: 2620) B*35: 01 HPVAEADYFEY EBV EBNA-1 407-417(4A) (SEQ ID NO: 2621) B*35: 01 HPNIEEVAL HCV NS3 1359-1367 (SEQ ID NO: 2622) B*35: 01 CPNSSIVY HCV E1 207-214 (SEQ ID NO: 2623) B*35: 01 NPDIVIYQY HIV-1 RT330-338 (SEQ ID NO: 2624) B*35: 01 NPDIVIYQY HIV-1 HIV-1 RT328-336 (SEQ ID NO: 2625) B*35: 01 VPLDEDFRKY HIV-1 HIV-1 RT273-282 (SEQ ID NO: 2626) B*40: 01 MEVTPSGTWL SARS-CoV-2 Nucleocapsid protein 322-330 (SEQ ID NO: 2627) (confirmed epitope) B*40: 01 GEARKTFVEL Dengue NS3 528-537 (SEQ ID NO: 2628) B*40: 01 IEDPPFNSL EBV LMP2 200-208 (SEQ ID NO: 2629) B*40: 01 REISVPAEIL HCV NS5a 2266-2275 (SEQ ID NO: 2630) B*40: 01 KEKGGLEGL HIV-1 Nef 92-100 (SEQ ID NO: 2631) C*06: 02 TRATKMQVI pp65 211-219 (SEQ ID NO: 2632) C*06: 02 QIKVRVDM 1E1 88-95 (SEQ ID NO: 2633) C*06: 02 TRRFLPQIL NS3 205-213 (SEQ ID NO: 2634) C*07: 02 CRVLCCYVL 1E1 309-317 (SEQ ID NO: 2635) E*01: 01 VMAPRTLIL HLA-C leader sequence peptide (SEQ ID NO: 2636) E*01: 01 VMAPRTLVL HLA-A leader sequence peptide (SEQ ID NO: 2637) DRB1*01: 01 DSVTPMILKAQKGGNL Dog dander Can f 1 33-48 (SEQ ID NO: 2641) DRB1*01: 01 KCIEWEKAQHGA Mugwort pollen Art v 1 25-36 (SEQ ID NO: 2642) DRB1*01: 01 LPVVLENARILKNCVDAK Cat dander Eel d 1 53-70 (SEQ ID NO: 2643) DRB1*01: 01 LRQMRTVTPIRMQGG House dust mite Der p1 96-110 (SEQ ID NO: 2644) DRB1*01: 01 VAAAPQVKYAVFEAALTKAI Timothy grass Phl p 5 p26 (SEQ ID NO: 2645) DRB1*01: 01 YESYKFIPALEAAVK Grass pollen allergen Phl p 5 196-210 (SEQ ID NO: 2646) DRB1*01: 01 ETLLRAVESYLLAHS Birch pollen allergen Bet v 1 142-156 (SEQ ID NO: 2647) DRB1*01: 01 ACYEFLWGPRALVETS MAGE-A3 267-282 (SEQ ID NO: 2648) DRB1*01: 01 LKEFTVSGNILTIRL NY-ESO-1 123-137 (SEQ ID NO: 2649) DRB1*01: 01 LLEFYLAMPFATPME NY-ESO-1 87-101 (SEQ ID NO: 2650) DRB1*01: 01 KVPIKWMALESILRRRF HER2 883-899 (SEQ ID NO: 2651) DRB1*01: 01 LPLKMLNIPSINVH CMV pp65 116-129 (SEQ ID NO: 2652) DRB1*01: 01 GAALQIPFAMQMAYRF SARS-CoV Sprotein (873-888) (SEQ ID NO: 2653) (conserved in SARS-CoV-2) DRB1*01: 01 MAYRFNGIGVTQNVLY SARS-CoV Sprotein (884-899) (SEQ ID NO: 2654) (conserved in SARS-CoV-2) DRB1*01: 01 QLIRAAEIRASANLAATK SARS-CoV Sprotein (993-1010) (SEQ ID NO: 2655) (conserved in SARS-CoV-2) DRB1*01: 01 TVFYNIPPMPL EBV EBNA2 280-290 (SEQ ID NO: 2656) DRB1*01: 01 TSLYNLRRGTALA EBV EBNA1 515-527 (SEQ ID NO: 2657) DRB1*01: 01 QAGFFLLTRILTIPQS HBV envelope 179-194 (SEQ ID NO: 2658) DRB1*01: 01 PPAYRPPNAPILSTL HBV core 158-172 (SEQ ID NO: 2659) DRB1*01: 01 LCWGELMTLATWVGVN HBV core 60-75 (SEQ ID NO: 2660) DRB1*01: 01 TLLFNILGGWVAA HCV polyprotein 1806-1818, NS4b (SEQ ID NO: 2661) DRB1*01: 01 DYVDRFYKTLRAE HIV-1 gag 295-307 (SEQ ID NO: 2662) DRB1*01: 01 KRWIILGLNKIVRMYSPTSI HIV-1 gag 263-282 (SEQ ID NO: 2663) DRB1*01: 01 FRDYVDRFYKTLRAEQASQE HIV-1 gag 293-312 (SEQ ID NO: 2664) DRB1*01: 01 SGPLKAEIAQRLEDV Influenza A MP 17-31 (SEQ ID NO: 2665) DRB1*01: 01 PKYVKQNTLKLAT Influenza A HA 307-319 (SEQ ID NO: 2666) DRB1*01: 01 EYLNKIQNSLSTEWSPCSVT CSP 326-345 (SEQ ID NO: 2667) DRB1*01: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*01: 01 QEIYMQHTYPISA TT257-269 (SEQ ID NO: 2669) DRB1*01: 01 GEEYLILSARDVLAV Mtb 10 kDa chaperonin GroES (SEQ ID NO: 2670) DRB1*03: 01 HTYTIDWTKDAVTWS Aspergillus fumigatus Crf1/p41 171-185 (SEQ ID NO: 2671) DRB1*03: 01 VYYLTRDPTTPLARAA HCV polyprotein 2800-2815, NS5b (SEQ ID NO: 2672) DRB1*03: 01 PIVQLQGDSNCLKCFR HPV E2 285-300 (SEQ ID NO: 2673) DRB1*03: 01 MEAIAKRLDACQDQLLELYE HPV E2 1-20 (SEQ ID NO: 2674) DRB1*03: 01 RQIFGDYKTTIC PLP 98-109 (SEQ ID NO: 2675) DRB1*03: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*03: 01 MAKTIAYDEEARR Mtb groEL 1-13 (SEQ ID NO: 2676) DRB1*03: 01 KTIAYDEEARR Mtb groEL 3-13 (SEQ ID NO: 2677) DRB1*04: 01 APYHFDLSGHAFG Rye grass Lol p1 124-136 (SEQ ID NO: 2678) DRB1*04: 01 ELEKYQQLNSERGVPN Cow dander Bos d 2 143-158 (SEQ ID NO: 2679) DRB1*04: 01 ETLLRAVESYLLAHS Birch pollen allergen Bet v 1 141-155 (SEQ ID NO: 2680) DRB1*04: 01 VAAAPQVKYAVFEAALTK Timothy grass Phl p 5 p26 AI (SEQ ID NO: 2645) DRB1*04: 01 FHTYTIDWTKDAVTW Aspergillus fumigatus Crf1/p41 170-184 (SEQ ID NO: 2681) DRB1*04: 01 NFIRMVISNPAAT GAD65 555-567 (SEQ ID NO: 2682) DRB1*04: 01 IAFTSEHSHFSLK GAD65 274-286 (SEQ ID NO: 2683) DRB1*04: 01 GAGSLQPLALEGSLQKRG Proinsulin 73-90 (SEQ ID NO: 2684) DRB1*04: 01 GIVEQCCTSICSLYQ Proinsulin 90-104 (SEQ ID NO: 2685) DRB1*04: 01 ATEG[cit]V[cit]VNS Cit aggregan 89-103 AYQDK (SEQ ID NO: 2686) DRB1*04: 01 ATIKAEFV[cit]AETPYM Cit CILP 297-311 (SEQ ID NO: 2687) DRB1*04: 01 DVMNILLQYVVKSFDRSTKV GAD65 113-132 (SEQ ID NO: 2030) DRB1*04: 01 GKLYGI[cit] Cit CILP 982-996 (SEQ ID NO: 2688) DRB1*04: 01 GVYAT[cit]SSAV[cit]L Cit vimentin aa 59-78 [cit]SSVPGVR (SEQ ID NO: 2689) DRB1*04: 01 IFDS[cit]GNPTVEVDLF Cit alpha-enolase 11-25 (SEQ ID NO: 2690) DRB1*04: 01 KGMAALPRLIAFTSEHSHFS GAD65 265-284 (SEQ ID NO: 2691) DRB1*04: 01 QDFTN[cit]INKLKNS Cit fibrinogen-alpha 79-91 (SEQ ID NO: 2692) DRB1*04: 01 WNRQLYPEWTEAQRLD gp100 44-59 (SEQ ID NO: 2693) DRB1*04: 01 ISPNSVFSQWRVVCDSLEDY Tyrp1 277-297 D (SEQ ID NO: 2694) DRB1*04: 01 ESEFQAALSRKVAKL MAGE-A6 102-116 (SEQ ID NO: 2695) DRB1*04: 01 LTQYFVQENYLEYRQVPG MAGE-A6 246-263 (SEQ ID NO: 2696) DRB1*04: 01 YACFVSNLATGRNNS CEA 653-667 (SEQ ID NO: 2697) DRB1*04: 01 NYTLRVDCTPLMYSL PSMA 459-473 (SEQ ID NO: 2698) DRB1*04: 01 IYRRRLMKQDFSVPQLPHS gp100 615-633 (SEQ ID NO: 2699) DRB1*04: 01 ALHIYMDGTMSQVQGSA tyrosinase 365-381 (SEQ ID NO: 2700) DRB1*04: 01 RNGYRALMDKSLHVGTQCA MART-1 51-73 LTRR (SEQ ID NO: 2701) DRB1*04: 01 LKEFTVSGNILTIRL NY-ESO-1 123-137 (SEQ ID NO: 2649) DRB1*04: 01 LLEFYLAMPFATPME NY-ESO-1 87-101 (SEQ ID NO: 2650) DRB1*04: 01 KVPIKWMALESILRRRF HER2 883-899 (SEQ ID NO: 2651) DRB1*04: 01 QALNTLVKQLSSNFGAI SARS-CoV Sprotein (939-955) (SEQ ID NO: 2702) (conserved in SARS-CoV-2) DRB1*04: 01 QLIRAAEIRASANLAATK SARS-CoV Sprotein (993-1010) (SEQ ID NO: 2655) (conserved in SARS-CoV-2) DRB1*04: 01 PYYVVDLSVRGM EBV BHRF1 122-133 (SEQ ID NO: 2703) DRB1*04: 01 AEGLRALLARSHVER EBV EBNA1 482-496 (SEQ ID NO: 2704) DRB1*04: 01 GQTYHLIVDTDSLGNPSLSV EBV EBNA2 11-30 (SEQ ID NO: 2705) DRB1*04: 01 LSFLPSDFFPSVRDL HBV core 45-59 (SEQ ID NO: 2706) DRB1*04: 01 GYKVLVLNPSVAATL HCV polyprotein 1248-1262, NS3 (SEQ ID NO: 2707) DRB1*04: 01 SGIQYLAGLSTLPGNPAIA HCV polyprotein 1770-1790, NS4b SL (SEQ ID NO: 2708) DRB1*04: 01 AFSPEVIPMFSALSEGATPQ HIV-1 gag 163-182 (SEQ ID NO: 2709) DRB1*04: 01 FWRGENGRKTRIAYERMCNI Influenza A NP 206-229 LKGK (SEQ ID NO: 2710) DRB1*04: 01 GFVFTLTVPSER Influenza A MP 61-72 (SEQ ID NO: 2711) DRB1*04: 01 PKYVKQNTLKLAT Influenza A HA 307-319 (SEQ ID NO: 2666) DRB1*04: 01 EYLNKIQNSLSTEWSPCSVT CSP 326-345 (SEQ ID NO: 2667) DRB1*04: 01 RTELLKDAIGEGK MOG 97-109 (SEQ ID NO: 2712) DRB1*04: 01 TWTTCQSIAFPSKTSASIGS PLP 180-199 (SEQ ID NO: 2713) DRB1*04: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*04: 01 SSPKPWIYATSNLAS Rituximab Light chain 41-55 (SEQ ID NO: 2714) DRB1*04: 02 PKYVKQNTLKLAT Influenza A HA 307-319 (SEQ ID NO: 2666) DRB1*04: 02 PVSKMRMATPLLMQA CLIP 87-101 321-3339 (SEQ ID NO: 2668) DRB1*04: 05 LWWVNNQSLPVSP CEA 331-3339 (SEQ ID NO: 2715) DRB1*04: 05 PKYVKQNTLKLAT Influenza A HA 307-319 (SEQ ID NO: 2666) DRB1*04: 05 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*07: 01 DRVNFKYSFSVIE Alder pollen Aln g 1 76-88 (SEQ ID NO: 2716) DRB1*07: 01 VAAAPQVKYAVFEAALTKAI Timothy grass Phl p 5 p26 (SEQ ID NO: 2645) DRB1*07: 01 VGLLKAVESYLLA Alder pollen Aln g 1 142-154 (SEQ ID NO: 2717) DRB1*07: 01 YYSNVTATRLLSSTNS Pertussis toxin subunit 2 129-144 (SEQ ID NO: 2718) DRB1*07: 01 EPDVYYTSAFVFPTK CMV pp65 177-191 (SEQ ID NO: 2719) DRB1*07: 01 PDDYSNTHSTRYVTV CMV gB 215-229 (SEQ ID NO: 2720) DRB1*07: 01 VPGLYSPCRAFFNKEELL EBV MCP 1264-1281 (SEQ ID NO: 2721) DRB1*07: 01 PGPLRESIVCYFMVFLOTHI EBV EBNA1 551-570 (SEQ ID NO: 2722) DRB1*07: 01 VYGGSKTSLYNLRRGTALAI EBV EBNA1 509-528 (SEQ ID NO: 2723) DRB1*07: 01 PRSPTVFYNIPPMPLPPSQL EBV EBNA2 276-295 (SEQ ID NO: 2724) DRB1*07: 01 AYCLWMMLLISQAEAALELI HCV N52 732-753 T (SEQ ID NO: 2725) DRB1*07: 01 WPLLLLLLALPQRAYAQ HCV N52 47-63 (SEQ ID NO: 2726) DRB1*07: 01 SLTITSLLRRHNWITSCS HCV NS5a 1957-1975 (SEQ ID NO: 2727) DRB1*07: 01 TTVRLRAYMNTPGLPVC HCV NS3 1535-1551 (SEQ ID NO: 2728) DRB1*07: 01 FRDYVDRFYKTLRAEQASQE HIV-1 gag 293-312 (SEQ ID NO: 2664) DRB1*07: 01 PKYVKQNTLKLAT Influenza A HA 307-319 (SEQ ID NO: 2666) DRB1*07: 01 EYLNKIQNSLSTEWSPCSVT CSP 326-345 (SEQ ID NO: 2667) DRB1*07: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*07: 01 WVKQTPGRGLEWIGA Rituximab Heavy chain 36-50 (SEQ ID NO: 2729) DRB1*07: 01 SSPKPWIYATSNLAS Rituximab Light chain 41-55 (SEQ ID NO: 2714) DRB1*07: 01 EWVAEIRSKSINSAT Infliximab Heavy chain 46-60 (SEQ ID NO: 2730) DRB1*09: 01 VAAAPQVKYAVFEAALTKAI Timothy grass Phl p 5 p26 (SEQ ID NO: 2645) DRB1*09: 01 LLEFYLAMPFATPME NY-ESO-1 87-101 (SEQ ID NO: 2650) DRB1*09: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*09: 01 MTEQQWNFAGIEAAA Mtb ESAT6 1-15 (SEQ ID NO: 2731) DRB1*11: 01 LEAAFNAEFNEIRRVLLEEN Peanut Ara h 1 321-340 (SEQ ID NO: 2732) DRB1*11: 01 TSRNNPFYFPSRRFSTRYGN Peanut Ara h 1 169-188 (SEQ ID NO: 2733) DRB1*11: 01 VVNKGTGNLELVAVRKEQQQ Peanut Ara h 1 457-476 (SEQ ID NO: 2734) DRB1*11: 01 VAAAPQVKYAVFEAALTKAI Timothy grass Phl p 5 p26 (SEQ ID NO: 2645) DRB1*11: 01 TSYVKVLHHMVKISG MAGE-A3 281-295 (SEQ ID NO: 2735) DRB1*11: 01 ENIQRFLPNPAGVQLEDPEF Factor VIII589-608 (SEQ ID NO: 2736) DRB1*11: 01 FRDYVDRFYKTLRAEQASQE HIV-1 gag 293-312 (SEQ ID NO: 2664) DRB1*11: 01 PKYVKQNTLKLAT Influenza A HA 307-319 (SEQ ID NO: 2666) DRB1*11: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*11: 01 VSIDKFRIFCKALNPK TT 1084-1099 (SEQ ID NO: 2737) DRB1*11: 01 FNNFTVSFWLRVPKVSASHL TT 947-967 E (SEQ ID NO: 2738) DRB1*11: 01 KFIIKRYTPNNEIDSF TT 1174-1189 (SEQ ID NO: 2739) DRB1*11: 01 QYIKANSKFIGITEL TT 830-844 (SEQ ID NO: 2740) DRB1*11: 01 WVKQTPGRGLEWIGA Rituximab Heavy chain 36-50 (SEQ ID NO: 2729) DRB1*15: 01 VAAAPQVKYAVFEAALTKAI Timothy grass Phl p 5 p26 (SEQ ID NO: 2645) DRB1*15: 01 VGLLKAVESYLLA Alder pollen Aln g 1 142-154 (SEQ ID NO: 2717) DRB1*15: 01 AVNIVGYSNAQGVDY House dust mite Der p1 251-265 (SEQ ID NO: 2741) DRB1*15: 01 ETLLRAVESYLLAHS Birch pollen allergen Bet v 1 142-156 (SEQ ID NO: 2647) DRB1*15: 01 LRQMRTVTPIRMQGG House dust mite Der p1 96-110 (SEQ ID NO: 2644) DRB1*15: 01 DENPVVHFFKNIVTPRTPP Myelin basic protein 83-101 (SEQ ID NO: 2027) DRB1*15: 01 MSIYVYALPLKMLNI CMV pp65 109-123 (SEQ ID NO: 2742) DRB1*15: 01 NFPYLVAYQATVCARA HCV polyprotein 1582-1597, NS3 (SEQ ID NO: 2743) DRB1*15: 01 GINAVAYYRGLDVSV HCV polyprotein 1411-1425, NS3 (SEQ ID NO: 2744) DRB1*15: 01 TTVRLRAYMNTPGLPVC HCV NS3 1535-1551 (SEQ ID NO: 2728) DRB1*15: 01 PVSKMRMATPLLMQA CLIP 87-101 (SEQ ID NO: 2668) DRB1*15: 01 SSPKPWIYATSNLAS Rituximab Light chain 41-55 (SEQ ID NO: 2714) DRB1*15: 01 EWVAEIRSKSINSAT Infliximab Heavy chain 46-60 (SEQ ID NO: 2730) DRB1*15: 01 MHVSFVMAYPEMLAA Mtb PEfamily protein (SEQ ID NO: 2745) DRB1*15: 01 MSQIMYNYPAMMAHA Mtb ESAT-6 like protein (SEQ ID NO: 2746)

As further illustrative and non-limiting examples, the following antigens provided in Table 6 are known in the art to be relevant to celiac disease. In some embodiments, the TCR recognition domain can comprise one or more of the peptides provided in Table 6. In some embodiments, the TCR recognition domain can comprise a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of the peptides of Table 6. In some embodiments, the TCR recognition domain can comprise a sequences with at least 95% sequence identity to one of peptides of Table 6, wherein that sequence retains the wild-type activity of the peptide of Table 6.

TABLE 6 The sequences are, in the order presented, SEQ ID NOs: 1000-2012. ID Type Description Toxicity Form HLADQ Refs SeqLen Sequence 1 alpha-gliadin alpha-gliadin CT-1 (p1-p22 of B 3142) Toxic Native Unknown 41 22 VPVPQLQPQNPSQQ QPQEQVPL 2 alpha-gliadin alpha-gliadin peptide CT-2 (p23-p53 of B 3142) Toxic Native Unknown 41 31 VQQQQFPGQQQPFP PQQPYPQPQPFPSQ QPY 3 alpha-gliadin alpha-gliadin p14 (p1-p19) Immunogenic Native DQ2 34 19 VRVPVPQLQPQNPS QQQPQ 4 alpha-gliadin alpha-gliadin p15 (p11-p28) Immunogenic Native DQ2 34 18 QNPSQQQPQEQVPL VQQQ 5 alpha-gliadin alpha-gliadin p209 Immunogenic Native DQ2 39, 34 20 FPGQQQPFPPQQPY PQPQPF 7 alpha-gliadin alpha-gliadin (p44-p55) Immuno- Native HLA-DR 10 12 PQPQPFPSQQPY genic, Toxic 8 alpha-gliadin Epitope DQ2-alpha-I/II/III Immunogenic Native DQ2 62 20 YLQLQPFPQPQLPY PQPQLP 9 alpha-gliadin alpha2-gliadin 1420 (p56-p70) Immunogenic Native DQ2, DQ8 17 15 YLQLQPFPQPQLPY P 10 alpha-gliadin alpha2-gliadin 33-mer (p57-p89) Immunogenic Native DQ2, DQ8 2, 25 33 LQLQPFPQPQLPYP (DQ2/8) QPQLPYPQPQLPYP QPQPF 11 alpha-gliadin Deamidated alpha2-gliadin 33-mer (p57-p89) Immunogenic Deamidated DQ2.5 61 33 LQLQPFPQPELPYP QPELPYPQPELPYP QPQPF 12 alpha-gliadin alpha-2 gliadin G8 (p56-p75) Toxic, Native DQ2 51, 63 20 LQLQPFPQPQLPYP Immunogenic QPQLPY 13 alpha-gliadin alpha-2 gliadin G9 (p56-p75; E65) Immunogenic Deamidated DQ2 51 20 LQLQPFPQPELPYP QPQLPY 14 alpha-gliadin alpha-9 gliadin G5 (p56-p68) Immunogenic Native DQ2 51 13 LQLQPFPQPQLPY 15 alpha-gliadin alpha-9 gliadin G5 (p56-p68; E65) Immunogenic Deamidated DQ2 51 13 LQLQPFPQPELPY 16 alpha-gliadin Wheat peptide W02 Immunogenic Native DQ2 62, 86 16 QLQPFPQPQLPYPQ PQ 17 alpha-gliadin Glia-alpha9 (p57-p71) Immunogenic Native DQ2  9 15 QLQPFPQPQLPYPQ P 18 alpha-gliadin Glia-alpha9 (p57-p71; T69 and H70) Immunogenic Native DQ2  9 15 QLQPFPQPQLPYTH P 19 alpha-gliadin Glia-alpha9 (p57-p71; R59) Immunogenic Native DQ2  9 15 QLRPFPQPQLPYPQ P 20 alpha-gliadin Glia-alpha9 (p57-p71; H63 and H70) Immunogenic Native DQ2  9 15 QLQPFPHPQLPYPH P 21 alpha-gliadin Glia-alpha9 (p57-p71; A63) Immunogenic Native DQ2  9 15 QLQPFPQAQLPYPQ P 22 alpha-gliadin Glia-alpha9 (p57-p70) Immunogenic Native DQ2  8 14 QLQPFPQPQLPYPQ 23 alpha-gliadin Glia-alpha9 (p57-p70; E65) Immunogenic Deamidated DQ2  8 14 QLQPFPQPELPYPQ 24 alpha-gliadin alpha-9 gliadin (p57-p68); alpha2/alpha9 gliadin Immunogenic Native DQ2 14, 23, 2, 12 QLQPFPQPQLPY 84 25 alpha-gliadin alpha-9 gliadin (p57-p68; E65); alpha-I Immunogenic Deamidated DQ2 43, 14, 8, 12 QLQPFPQPELPY 84 26 alpha-gliadin alpha-9 gliadin epitope homolog (p57-p68; E63 Immunogenic Native DQ2  8 12 QLQPFPEPQLPY (considered native form of synthetic)) 27 alpha-gliadin alpha-9 gliadin epitope homolog (p57-p68; E63 and Immunogenic Deamidated DQ2  8 12 QLQPFPEPELPY E65 (tTG-treated form)) 28 alpha-gliadin alpha-9 gliadin epitope homolog (p57-p68; Q64 Immunogenic Native DQ2  8 12 QLQPFPQQQLPY (considered native form of synthetic)) 29 alpha-gliadin alpha-9 gliadin epitope homolog (p57-p68; Q64 and Immunogenic Deamidated DQ2  8 12 QLQPFPEQQLPY E63 (tTG-treated form)) 30 alpha-gliadin alpha-9 gliadin epitope homolog (p57-p68; Q64 and Immunogenic Deamidated DQ2  8 12 QLQPFPQQELPY E65 (tTG-treated form)) 31 alpha-gliadin alpha-9 gliadin epitope homolog (p57-p68; Q64, E63 Immunogenic Deamidated DQ2  8 12 QLQPFPEQELPY and E65 (tTG-treated form)) 32 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72) Immunogenic Native DQ2 59 15 LQPFPQPQLPYPQP Q 33 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; E65) Immunogenic Deamidated DQ2 59 15 LQPFPQPELPYPQP Q 34 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; S64) Immunogenic Native DQ2 59 15 LQPFPQSQLPYPQP Q 35 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; S64 and E65) Immunogenic Deamidated DQ2 59 15 LQPFPQSELPYPQP Q 36 alpha-gliadin DQ2-alpha-I epitope (p58-p68) Immunogenic Native DQ2 47 11 LQPFPQPQLPY 37 alpha-gliadin DQ2-alpha-I epitope (p58-p68; E65 considered Immunogenic Deamidated DQ2 47 11 LQPFPQPELPY Deamidated form) 38 alpha-gliadin Glia-alpha2 (p60-p76) Immunogenic Native DQ2  8 17 QPFPQPQLPYPQPQ LPY 39 alpha-gliadin Glia-alpha2 (p60-p76; E66) Immunogenic Deamidated DQ2  8 17 QPFPQPELPYPQPQ LPY 40 alpha-gliadin Glia-alpha2 (p60-p76; E73) Immunogenic Deamidated DQ2  8 17 QPFPQPQLPYPQPE LPY 41 alpha-gliadin Glia-alpha2 (p60-p76; E66 and E73) Immunogenic Deamidated DQ2  8 17 QPFPQPELPYPQPE LPY 42 alpha-gliadin Glia-alpha2 (p60-p69) Immunogenic Native DQ2 45 10 QPFPQPQLPY 43 alpha-gliadin Glia-alpha2 (p60-p69; E66) Immunogenic Deamidated DQ2 45 10 QPFPQPELPY 44 alpha-gliadin alpha2-gliadin 1421 (p61-p75) Immunogenic Native DQ2, DQ8 17 15 PFPQPQLPYPQPQL P 45 alpha-gliadin Glia-alpha2 (p61-p71) Immunogenic Native DQ2 59 11 PFPQPQLPYPQ 46 alpha-gliadin Glia-alpha2 (p61-p71; E66) Immunogenic Deamidated DQ2 59 11 PFPQPELPYPQ 47 alpha-gliadin Glia-alpha2 (p61-p71; T70 and H71) Immunogenic Native DQ2 59 11 PFPQPQLPYTH 48 alpha-gliadin Glia-alpha2 (p61-p71; T70, H71 and E66) Immunogenic Deamidated DQ2 59 11 PFPQPELPYTH 49 alpha-gliadin Glia-alpha2 (p61-p71; H64) Immunogenic Native DQ2 59 11 PFPHPQLPYPQ 50 alpha-gliadin Glia-alpha2 (p61-p71; H64 and E66) Immunogenic Deamidated DQ2 59 11 PFPHPELPYPQ 53 alpha-9 gliadin DQ2.5_glia_alpha-la Immunogenic Native DQ2.5 14, 90, 17, 9 PFPQPQLPY 23 54 alpha-9 gliadin DQ2.5_glia_alpha-la Immunogenic Deamidated DQ2.5 14, 90, 17, 9 PFPQPELPY 23 55 alpha-gliadin alpha-2 gliadin 1206 (p62-p75) Immunogenic Native DQ2 14, 1, 2 14 PQPQLPYPQPQLPY 56 alpha-gliadin alpha-II/alpha-III epitope (p62-p75; E72) Immunogenic Deamidated DQ2 14 14 PQPQLPYPQPELPY 57 alpha-gliadin alpha-2 gliadin (p62-p75; E65) Immunogenic Deamidated DQ2 14, 25 14 PQPELPYPQPQLPY 58 alpha-gliadin alpha-2 gliadin (p62-p75; E65 and E72) Immunogenic Deamidated DQ2 14 14 PQPELPYPQPELPY 59 alpha-gliadin G4-9A gliadin (p62-p75; E65 and A70) Immunogenic Deamidated DQ2 51 14 PQPELPYPAPQLPY 60 alpha-gliadin G4-11A gliadin (p62-p75; E65 and A72) Immunogenic Deamidated DQ2 51 14 PQPELPYPQPALPY 61 alpha-gliadin gliadin (p62-p75; E65 and A73) Immunogenic Deamidated DQ2 51 14 PQPELPYPQPQAPY 62 alpha-gliadin gliadin (p62-p75; E65 and A74) Immunogenic Deamidated DQ2 51 14 PQPELPYPQPQLAY 63 alpha-gliadin gliadin (p62-p75; E65 and A70) Immunogenic Deamidated DQ2 51 14 PQPELPYPQPQLPA 64 alpha-gliadin alpha-2 gliadin (p62-p73) Immunogenic Native DQ2 25, 47 12 PQPQLPYPQPQL 65 alpha-gliadin alpha-2 gliadin (p62-p73; E65) Immunogenic Deamidated DQ2 25, 47 12 PQPELPYPQPQL 66 alpha-gliadin alpha-ll (p62-p72) Immunogenic Native DQ2 43 11 PQPQLPYPQPQ 67 alpha-gliadin alpha-ll (p62-p72; E65 and E72) Immunogenic Deamidated DQ2 43 11 PQPELPYPQPE 68 alpha-2 gliadin CAUTION 100% match to one fungal protein and many  Immunogenic Native DQ2.5 14, 17, 23 9 PQPQLPYPQ wheat proteins with multiple epitopes DQ2.5_glia_alpha2 69 alpha-2 gliadin DQ2.5_glia alpha2 Immunogenic Deamidated DQ2.5 14, 17, 23 9 PQPELPYPQ 70 alpha-gliadin alpha2-gliadin 1423 (p71-p85) Immunogenic Native DQ2, DQ8 17 15 QPQLPYPQPQLPYP Q 71 alpha-gliadin a-gliadin (p62-p84) Immunogenic Native DQ2 62 20 PQLPYPQPQLPYPQ PQLPYP 72 alpha-gliadin alpha-Ill-gliadin (p62-p79) Immunogenic Native DQ2 25 12 PQLPYPQPQLPY 73 alpha-gliadin alpha-Ill-gliadin (p62-p79; E76) Immunogenic Deamidated DQ2 25 12 PQLPYPQPELPY 74 alpha-gliadin alpha-I epitope Immunogenic Native DQ2 25 12 LQLPFPQPQLPY 75 alpha-gliadin alpha-I epitope Deamidated form Immunogenic Deamidated DQ2 25 12 LQLPFPQPELPY 76 alpha-gliadin Glia-alpha2 25-mer (p64-p89) Immunogenic Native DQ2 57 25 PQLPQFLQPQPYPQ PQLPYPQPQPF 77 alpha-gliadin Glia-alpha2 25-mer (p64-p89; E65) Immunogenic Deamidated DQ2 57 25 PELPQFLQPQPYPQ PQLPYPQPQPF 78 alpha-gliadin Glia-alpha2 25-mer (p64-p89; E79) Immunogenic Deamidated DQ2 57 25 PQLPQFLQPQPYPQ PELPYPQPQPF 79 alpha-gliadin Glia-alpha2 25-mer (p64-p89; E65 and E79) Immunogenic Deamidated DQ2 57 25 PELPQFLQPQPYPQ PELPYPQPQPF 80 alpha-gliadin alpha2-gliadin 1422 (p66-p80) Immunogenic Native DQ2 17 15 QLPYPQPQLPYPQP Q 81 alpha-gliadin alpha-III epitope (p66-p78) Immunogenic Native DQ2 47 13 QLPYPQPQLPYPQ 82 alpha-gliadin alpha-III epitope (p66-p78; E72) Immunogenic Deamidated DQ2 47 13 QLPYPQPELPYPQ 83 alpha-gliadin Wheat peptide W01 Immunogenic Native DQ2 62 12 LPYPQPQLPYPQ 84 alpha-3 gliadin DQ2.5_glia_alpha 1b Immunogenic Native DQ2.5 23 9 PYPQPQLPY 85 alpha-3 gliadin DQ2.5_glia_alpha 1b Immunogenic Deamidated DQ2.5 23 9 PYPQPELPY 86 alpha-gliadin Glia-alpha2 18-mer (p71-p89) Immunogenic Native DQ2 82 18 QPQPYPQPQLPYPQ PQPF 87 alpha-gliadin Glia-alpha2 18-mer (p71-p89; E79) Immunogenic Deamidated DQ2 82, 57 18 QPQPYPQPELPYPQ PQPF 88 alpha-gliadin Wheat peptide W18 Immunogenic Native DQ2 62, 82 20 PQLPYPQPQLPYPQ PQPFRP 89 alpha-gliadin Wheat peptide W18 Immunogenic Deamidated DQ2 62 20 PQLPYPQPELPYPQ PQPFRP 90 alpha-gliadin Wheat peptide W18 Immunogenic Native DQ2 62 16 YPQPQLPYPQPQPF RP 91 alpha-gliadin Wheat peptide W18 Immunogenic Deamidated DQ2 62 16 YPQPELPYPQPQPF RP 92 alpha-gliadin alpha2-gliadin 1424 (p76-p90) Immunogenic Native DQ2 17 15 YPQPQLPYPQPQPF R 93 alpha-20 DQ2.5_glia_alpha 3 Immunogenic Native DQ2.5 90, 8 9 FRPQQPYPQ gliadin 94 alpha-20 DQ2.5_glia_alpha 3 Immunogenic Deamidated DQ2.5 90, 8 9 FRPEQPYPQ gliadin 95 alpha-gliadin Gliadin (p198-p222) Immunogenic Native DQ8 (DQ2/8,  3 25 QQPQQQYPSGQGSF DQ1/8) QPSQQNPQAQG 96 alpha-gliadin alpha-2 gliadin (p219-p242) AJ133612 Immunogenic Native DQ8  6 24 QQPQQQYPSGQGSF QPSQQNPQAQ 97 alpha-gliadin alpha-2 gliadin (p219-p242; E229 and E237) AJ133612 Immunogenic Deamidated DQ8  6 24 QQPQQQYPSGEGSF QPSQENPQAQ 98 alpha-gliadin alpha-2 gliadin (p219-p242; E229) AJ133612 Immunogenic Deamidated DQ8  6 24 QQPQQQYPSGEGSF QPSQQNPQAQ 99 alpha-gliadin alpha-2 gliadin (p219-p242; E237) AJ133612 Immunogenic Deamidated DQ8  6 24 QQPQQQYPSGQGSF QPSQENPQAQ 100 alpha-gliadin alpha-gliadin (p220-p239) P18573 Immunogenic Native DQ8 54 20 QPQQQYPSGQGSFQ PSQQNP 101 alpha-gliadin Gda09 (p202-p219) P18573 Immunogenic Native DQ8 (DQ2/8, 3, 4 18 QQYPSGQGSFQPSQ DQ1/8) QNPQ 102 alpha-gliadin Gda09 (p203-p220) P18573 (alpha-gliadin (alpha-I)) Immunogenic Native DQ8  5 18 QYPSGQGSFQPSQQ NPQA 103 alpha-gliadin Gda09 (p203-p220; E216) P18573 (alpha-gliadin Immunogenic Deamidated DQ8  5 18 QYPSGEGSFQPSQE (alpha-l) NPQA 104 alpha-gliadin alpha2-gliadin 1447 (p226-p240) Immunogenic Native DQ8 17 15 YPSGQGSFQPSQQN P 105 alpha-gliadin Gliadin (p205-p222) Immunogenic Native DQ8 (DQ2/8)  3 18 PSGQGSFQPSQQNP QAQG 106 alpha-gliadin Gliadin (p205-p216); alpha2-gliadin Immunogenic Native DQ8 (DQ2/8) 46, 3 12 PSGQGSFQPSQQ 107 alpha-gliadin Gliadin (p205-p215); alpha2-gliadin Immunogenic Native DQ8 (DQ2/8) 46, 3 11 PSGQGSFQPSQ 108 alpha-gliadin Gda09 (p206-p217) P18573 Immunogenic Native DQ8 (DQ2/8) 46, 4 12 SGQGSFQPSQQN 109 alpha-gliadin Gda09 (p206-p217; E215) P18573 Immunogenic Deamidated DQ8 (DQ2/8) 46 12 SGQGSFQPSEQN 110 alpha-gliadin Gda09 (p206-p217; E208) P18573 Immunogenic Deamidated DQ8 (DQ2/8) 46, 83, 4 12 SGEGSFQPSQQN 111 alpha-gliadin Gda09 (p206-p217; E216) P18573 Immunogenic Deamidated DQ8 (DQ2/8) 46, 4 12 SGQGSFQPSQEN 112 alpha-gliadin Gda09 (p206-p217; E208 and E216) P18573 Immunogenic Deamidated DQ8 (DQ2/8) 3, 4 12 SGEGSFQPSQEN 113 alpha-gliadin alpha2-gliadin (p228-p236) Immunogenic Native DQ8 (DQ2/8)  6 9 GQGSFQPSQ 115 alpha-2 gliadin CAUTION 100% identity match to Dicot plant protein Immunogenic Native DQ8 (DQ2/8) 6, 90, 3 9 QGSFQPSQQ DQ8.5_glia_alpha 1 and many wheat family DQ8_glia_alpha 1 116 alpha-2 gliadin DQ8_glia_alpha 1 DQ8.5_glia_alpha 1 Immunogenic Deamidated DQ8 (DQ2/8) 90, 17, 83 9 EGSFQPSQQ 117 alpha-2 gliadin DQ8_glia_alpha 1 DQ8.5_glia_alpha 1 Immunogenic Deamidated DQ8 (DQ2/8) 90, 17 9 QGSFQPSQE 118 alpha-2 gliadin DQ8_glia_alpha 1 DQ8.5_glia_alpha 1 Immunogenic Deamidated DQ8 (DQ2/8) 90, 17 9 EGSFQPSQE 119 alpha-gliadin alpha2-gliadin 1448 (p231-p245) Immunogenic Native DQ8 17 15 GSFQPSQQNPQAQG S 120 alpha-gliadin alpha2-gliadin 1450 (p241-p255) Immunogenic Native DQ8 and 17 15 QAQGSVQPQQLPQF weak DQ2 E 121 alpha-gliadin Wheat peptide W02 Immunogenic Native DQ2 62 20 MQLQPFPQPQLPYP QPQLPY 122 alpha-gliadin Wheat peptide W02 Immunogenic Deamidated DQ2 62 20 MQLQPFPQPELPYP QPQLPY 123 alpha-gliadin Wheat peptide W02 Immunogenic Deamidated DQ2 62 16 QLQPFPQPELPYPQ PQ 124 alpha-gliadin Wheat peptide W02 Immunogenic Deamidated DQ2 62 16 ELQPFPQPELPYPQ PQ 125 alpha-gliadin Wheat peptide W02 Immunogenic Native DQ2 62 12 QPFPQPQLPYPQ 126 alpha-gliadin Wheat peptide W02 Immunogenic Deamidated DQ2 62 12 QPFPQPELPYPQ 127 gamma-gliadin Wheat peptide W01 Immunogenic Native DQ2 62 20 PQPFPPQLPYPQPQ LPYPQP 128 gamma-gliadin Wheat peptide W01 Immunogenic Deamidated DQ2 62 20 PQPFPPQLPYPQPE LPYPQP 129 gamma-gliadin Wheat peptide W01 Immunogenic Deamidated DQ2 62 12 LPYPQPELPYPQ 130 alpha-gliadin Wheat peptide W34 Immunogenic Native DQ2 62 20 VAHAIIMHQQQQQQ QEQKQQ 131 alpha-gliadin Wheat peptide W34 Immunogenic Native DQ2 62 16 VAHAIIMHQQQQQQ QE 132 alpha-gliadin Analog of alpha-gliadin (p31-p49; A31) Toxic Native DQ2 50 19 AGQQQPFPPQQPYP QPQPF 133 alpha-gliadin Analog of alpha-gliadin (p31-p49; A36) Toxic Native DQ2 50 19 LGQQQAFPPQQPYP QPQPF 134 alpha-gliadin alpha20-gliadin (p91-p106) Immunogenic Native DQ2  8 16 PQPFRPQQPYPQPQ PQ 135 alpha-gliadin alpha20-gliadin (p93-106; E97) Immunogenic Deamidated DQ2  8 16 PQPFRPEQPYPQPQ PQ 136 alpha-gliadin Glia-alpha20-gliadin (p93-p106) Immunogenic Native DQ2 19 14 PFRPQQPYPQPQPQ 137 alpha-gliadin Glia-alpha20-gliadin (p93-p106; E97) Immunogenic Deamidated DQ2 19 14 PFRPEQPYPQPQPQ 138 alpha-gliadin Glia-alpha20-gliadin (p96-106) minimal epitope Immunogenic Native DQ2 19 11 PQQPYPQPQPQ 139 alpha-gliadin  Glia-alpha20-gliadin (p96-106; E97) minimal epitope, Immunogenic Deamidated DQ2 19 11 PEQPYPQPQPQ synthetic 140 alpha-gliadin alpha-gliadin(p123-p132) Immunogenic Native DQ8 64, 30 10 QUPCMDVVL 141 alpha-gliadin alpha-gliadin (p206-p217) Toxic Native DQ2(Al B8 35 12 LGQGSFRPSQQN DR3 DQ2 and A24 B8 DR3 13 DQ2) 142 alpha-gliadin Peptide XT (1-55) Toxic Native Unknown 29 55 VRVPVPQLQPQNPS QQQPQEQVPLVQQQ QFLGQQQPFPPQQP YPQPQPFPSQQPY 143 alpha-gliadin Peptide XT (pl-p30) Toxic Native Unknown 29 30 VRVPVPQLQPQNPS QQQPQEQVPLVQQQ QF 144 alpha-gliadin alpha-gliadin B 3142 (p3-p55) Immunogenic, Native Unknown 40 53 VPVPQLQPQNPSQQ Toxic QPQEQVPLVQQQQF GGQQQPFPPQQPYP QPQPFPSQQPY 146 alpha-gliadin alpha-gliadin p19 (p21-p40) Immunogenic Native DQ2 34 20 QVPLVQQQQFLGQQ QPFPPQ 147 alpha-gliadin alpha-gliadin p134 Immunogenic Native DQ2 39 19 QFLGQQQPFPPQQP (alpha1*0501, YPQPQ 131*0201) 148 alpha-gliadin alpha-gliadin p135 Immunogenic Native DQ2 39 18 FLGQQQPFPPQQPY (alpha1*0501, PQPQ 131*0201) 149 alpha-gliadin Peptide XT (p31-p55) Immunogenic, Native Unknown 10, 29 25 LGQQQPFPPQQPYP Toxic QPQPFPSQQPY 150 alpha-gliadin alpha-gliadin (p31-p49) Immunogenic, Native DQ2 39, 11, 49, 19 LGQQQPFPPQQPYP Toxic (alpha1*0501, 34 QPQPF 131*0201) 151 alpha-gliadin alpha-gliadin p126 Immunogenic Native DQ2 10, 39, 13, 17 LGQQQPFPPQQPYP (alpha1*0501, 15, 66, 79 QPQ 131*0201) 152 alpha-gliadin alpha-gliadin (p31-p43) Immunogenic, Native HLA-DR 10, 13, 15, 13 LGQQQPFPPQQPY Toxic 72, 79 153 alpha-gliadin alpha-gliadin CAB76960 (p253-p272) Immunogenic Native DQ8 54 20 AMCNVYIPPYCAMA PFGIFG 154 alpha-gliadin alpha-gliadin (proline-rich domain) Immunogenic Native Unknown 42 16 CPQPFPSQQPYLQL QG 155 alpha-gliadin alpha-gliadin (p5-p22) (proline-rich domain) Immunogenic Native Unknown 42 18 CPQLQPQNPSQQQP QEQG 156 alpha-gliadin alpha-gliadin (p51-p70) Toxic Native DQ2 37 20 SQQPYLQLQPFPQP QLPYSQ 157 alpha-gliadin Wheat peptide W08 Immunogenic Native DQ2 62 20 LQLQPFPQPQLPYS QPQPFR 158 alpha-gliadin Wheat peptide W08 Immunogenic Deamidated DQ2 62 20 LQLQPFPQPELPYS QPQPFR 159 alpha-gliadin Glia-alpha9 (p57-p71; S69) Immunogenic Native DQ2  9 15 QLQPFPQPQLPYSQ P 160 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; S69) Immunogenic Native DQ2 59 15 LQPFPQPQLPYSQP Q 161 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; S69 and E65) Immunogenic Deamidated DQ2 59 15 LQPFPQPELPYSQP Q 162 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; S64 and S69) Immunogenic Native DQ2 59 15 LQPFPQSQLPYSQP Q 163 alpha-gliadin DQ2-Glia-alpha1 epitope (p58-p72; S64, S69 and E65) Immunogenic Deamidated DQ2 59 15 LQPFPQSELPYSQP Q 164 alpha-gliadin Wheat peptide W08 Immunogenic Native DQ2 62 12 QPFPQPQLPYSQ 165 alpha-gliadin Wheat peptide W08 Immunogenic Deamidated DQ2 62 12 QPFPQPELPYSQ 166 alpha-gliadin Glia-alpha Immunogenic Native DQ2 59 11 PFPQPQLPYSQ 167 alpha-gliadin Glia-alpha in Deamidated form Immunogenic Deamidated DQ2 59 11 PFPQPELPYSQ 168 alpha-gliadin alpha-gliadin (p202-p220) Toxic Native DQ2 11 19 QQYPLGQGSFRPSQ QNPQA 169 alpha-gliadin alpha-gliadin CAB76961 (p251-p270) Immunogenic Native DQ8 54 20 VYIPPYCTIAPFGI FGTNYR 170 alpha-gliadin Wheat peptide W13 Immunogenic Native DQ2 62 20 LQLQPFPQPQLPYL QPQPFR 171 alpha-gliadin Wheat peptide W13 Immunogenic Deamidated DQ2 62 20 LQLQPFPQPELPYL QPQPFR 172 alpha-gliadin Glia-alpha9 (p57-p71; L69) Immunogenic Native DQ2  9 15 QLQPFPQPQLPYLQ P 173 alpha-gliadin Wheat peptide W13 Immunogenic Native DQ2 62 12 QPFPQPQLPYLQ 174 alpha-gliadin Wheat peptide W13 Immunogenic Deamidated DQ2 62 12 QPFPQPELPYLQ 175 alpha-gliadin alpha-gliadin 4037 Immunogenic Native Unknown 60 17 PPYCTIVPFGIFGT NYR 176 alpha-gliadin alpha-gliadin Immunogenic Native DQ2 62 20 LQLQPFPQPQLPYP QPQPFR 177 alpha-gliadin alpha-Glia (p57-p73) Immunogenic Native DQ2 57 17 QLQPFPQPQLPYPQ PQP 178 alpha-gliadin alpha-Glia (p57-p73; E65) Immunogenic Deamidated DQ2 57 17 QLQPFPQPELPYPQ PQP 179 alpha-gliadin alpha-Glia (p57-p73; T65 and S73) Immunogenic Native DQ2 57 17 QLQPFPQPTLPYPQ PQS 180 alpha-gliadin alpha-gliadin (p57-p73; S73) Immunogenic Native DQ2 28 17 QLQPFPQPQLPYPQ PQS 181 alpha-gliadin alpha-gliadin (p57-p73; S73 and E65) Immunogenic Deamidated DQ2 28 17 QLQPFPQPELPYPQ PQS 182 alpha-gliadin Wheat peptide W09 Immunogenic Native DQ2 62 20 LQPFPQPQPFLPQL PYPQPQ 183 alpha-gliadin alpha-Glia AG11 (p78-p95) Immunogenic Native DQ2 57 17 PQPQPFLPQLPYPQ PQS 184 alpha-gliadin alpha-Glia AG11 (p78-p95; E86) Immunogenic Deamidated DQ2 57 17 PQPQPFLPELPYPQ PQS 185 alpha-gliadin Wheat peptide W09 Immunogenic Native DQ2 62 14 QPQPFLPQLPYPQP 186 alpha-gliadin Wheat peptide W09 Immunogenic Deamidated DQ2 62 14 EPQPFLPELPYPQP 187 alpha-gliadin Wheat peptide W09 Immunogenic Native DQ2 62 12 PQPFLPQLPYPQ 188 alpha-gliadin alpha-gliadin p211 Immunogenic Native DQ2 34 20 FPGQQQQFPPQQPY PQPQPF 189 alpha-gliadin alpha-Glia AG12 (p82-p98) Immunogenic Native DQ2 57 17 PQPQPFPPQLPYPQ PQS 190 alpha-gliadin alpha-Glia AG12 (p82-p98; E90) Immunogenic Deamidated DQ2 57 17 PQPQPFPPELPYPQ PQS 191 omega-gliadin Gliadin AAG17702 (p80-p99) Immunogenic Native DQ8 54 20 PFTQPQQPTPIQPQ QPFPQQ 192 omega-gliadin Wheat peptide W27 Immunogenic Native DQ2 62 11 PFTQPQQPTPI 193 omega-gliadin Gliadin AAG17702 (p88-p107) Immunogenic Native DQ8 54 20 TPIQPQQPFPQQPQ QPQQPF 194 omega-gliadin Wheat peptide W25 Immunogenic Native DQ2 62 11 TPIQPQQPFPQ 195 omega-gliadin Wheat peptide W30; Rye peptide R28 Immunogenic Native DQ2 62 12 PQQPFPQQPQQP 197 omega-gliadin omega-gliadin Immunogenic Native DQ2 62 20 PQQPQQPQQPFPQP QQPFPW 198 omega-gliadin Epitope DQ2-omega-I/II Immunogenic Native DQ2 62 20 PQQPQQPFPQPQQP FPWQPQ 199 omega-gliadin p4-p18 omega-gliadin of AAG17702 (p81-p102) Immunogenic Native DQ2 62 15 PQQPQQPFPQPQQP F 200 omega-gliadin p5-p19 omega-gliadin of AAG17702 (p81-p102) Immunogenic Native DQ2 62 15 QQPQQPFPQPQQPF P 201 omega-gliadin DQ2-omega-1 omega-Glia (p102-p118) Immunogenic Native DQ2 57 17 QPQQPFPQPQQPFP WQP 202 omega-gliadin DQ2-omega-1 omega-Glia (p102-p118; E104) Immunogenic Deamidated DQ2 57 17 QPEQPFPQPQQPFP WQP 203 omega-gliadin Wheat peptide W03, W19, B01 Immunogenic Deamidated DQ2 62, 86 17 QPEQPFPQPEQPFP WQP 204 omega-gliadin omega-Glia 17mer Immunogenic Deamidated DQ2.5 61 17 QPQQPFPQPEQPFP WQP 205 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 KPFPQPEQPFPWQP (p89-p102; E95 K89) 206 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 QKFPQPEQPFPWQP (p89-p102; E95 K90) 207 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 QPKPQPEQPFPWQP (p89-p102; E95 K91) 208 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 QPFPQPEQPFKWQP (p89-p102; E95 K99) 209 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 QPFPQPEQPFPKQP (p89-p102; E95 K100) 210 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 QPFPQPEQPFPWKP (p89-p102; E95 K101) 211 omega-gliadin omega-gliadin AAG17702 substituted by Lysine Immunogenic Native DQ2 62 14 QPFPQPEQPFPWQK (p89-p102; E95 K102) 212 omega-gliadin p6-p20 omega-gliadin of AAG17702 (p81-p102) Immunogenic Native DQ2 62 15 QPQQPFPQPQQPFP W 213 omega-gliadin p7-p21 omega-gliadin of AAG17702 (p81-p102) Immunogenic Native DQ2 62 15 PQQPFPQPQQPFPW Q 214 omega-gliadin p8-p22 omega-gliadin of AAG17702 (p81-p102), Immunogenic Native DQ2 62 15 QQPFPQPQQPFPWQ Wheat peptide W3 P 215 omega-gliadin Wheat peptide W03 Immunogenic Deamidated DQ2 62 15 EQPFPQPEQPFPWQ P 216 omega-gliadin Wheat peptide W03, W19, Barley peptide B01 Immunogenic Native DQ2 62 20 QPFPQPQQPFPWQP QQPFPQ 217 omega-gliadin Wheat peptide W03, W19, Barley peptide B01 Immunogenic Deamidated DQ2 62 20 QPFPQPEQPFPWQP QQPFPQ 218 omega-gliadin Wheat peptide W03, Barley peptide B01 Immunogenic Native DQ2 62 12 QPFPQPQQPFPW 219 omega-gliadin Wheat peptide W03, Barley peptide B01 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPFPW 220 omega-II gliadinDQ2.5_glia_omega 2 Immunogenic Deamidated DQ2.5 88, 90, 62 9 PQPEQPFPW 221 omega-II gliadinDQ2.5_glia_omega 2 Immunogenic Native DQ2 88, 90, 62 9 PQPQQPFPW 222 omega-gliadin Wheat peptide W19, Barley peptide B19 Immunogenic Native DQ2 62 12 PFPWQPQQPFPQ 223 omega-gliadin Wheat peptide W19 Immunogenic Deamidated DQ2 62 12 PFPWQPEQPFPQ 224 omega-gliadin Wheat peptide W30 Immunogenic Native DQ2 62 20 PLQPQQPFPQQPQQ PFPQPQ 225 omega-gliadin omega-gliadin Immunogenic Native DQ2 62 20 FPQQPQQPFPQPQL PFPQQS 226 omega-gliadin Wheat peptide W06 Immunogenic Native DQ2 62 20 QQPQQPFPQPQLPF PQQSEQ 227 omega-gliadin Wheat peptide W06 Immunogenic Native DQ2 62 12 QPFPQPQLPFPQ 228 omega-gliadin Gliadin AAG17702 p173-p192 Immunogenic Native DQ8 54 20 QQPFPQQPQQPFPQ PQQPIP 229 omega-gliadin Wheat peptide W32, Barley peptide B25, Rye peptide Immunogenic Native DQ2 62 12 PFPQQPQQPFPQ R26 230 omega-gliadin Wheat peptide W04 Immunogenic Native DQ2 62 20 PQQPQQPFPQPQQP IPVQPQ 231 omega-gliadin Wheat peptide W04 Immunogenic Native DQ2 62 11 PFPQPQQPIPV 232 omega-gliadin Gliadin AAG17702 p186-p205 Immunogenic Native DQ8 54 20 QPQQPIPVQPQQSF PQQSQQ 233 omega-gliadin Wheat peptide W20 Immunogenic Native DQ2 62 20 FPELQQPIPQQPQQ PFPLQP 234 omega-gliadin Wheat peptide W20 Immunogenic Native DQ2 62 12 PIPQQPQQPFPL 235 omega-gliadin Gliadin AAG17702 p225-p244 Immunogenic Native DQ8 54 20 PQQPQQPFPLQPQQ PFPQQP 236 omega-gliadin Wheat peptide W26, Barley peptide B20 Immunogenic Native DQ2 62 12 PFPLQPQQPFPQ 237 omega-gliadin Gliadin AAG17702 p239-p258 Immunogenic Native DQ8 54 20 PFPQQPQQPFPQQP QQSFPQ 246 omega5-liadin/ Glu-5 peptide epitope in native form Immunogenic Native DQ2 21 12 QQQQIPQQPQQF LMW glutenin 247 omega5-liadin/ Glu-5 peptide epitope in native form Immunogenic Native DQ2 21 12 QQQQLPQQPQQF LMW glutenin 248 omega5-liadin/ Glu-5 peptide epitope in Deamidated form Immunogenic Deamidated DQ2 21 12 QEQQIPEQPQQF LMW glutenin 249 omega5-liadin/ Glu-5 peptide epitope in Deamidated form Immunogenic Deamidated DQ2 21 12 QEQQLPEQPQQF LMW glutenin 252 omega5-liadin/  CAUTION 100% matches with 4 fungal proteins but Immunogenic Native DQ2 19 9 QIPQQPQQF LMW glutenin multiple epitopes on Triticum Glu-5 minimal epitope in native form 253 omega5-liadin/ Glu-5 minimal epitope in Deamidated form Immunogenic Deamidated DQ2 19 9 QIPEQPQQF LMW glutenin 254 omega5-liadin/ CAUTION 100% match with fungal and parasite proteins, Immunogenic Native DQ2 19 9 QLPQQPQQF LMW glutenin less with Glu-5 minimal epitope in native form 255 omega5-liadin/ Glu-5 minimal epitope in Deamidated form Immunogenic Deamidated DQ2 19 9 QLPEQPQQF LMW glutenin 256 omega5-liadin/ Glu-5 minimal epitope in Deamidated form Immunogenic Deamidated DQ2 (DQ2.2 44 9 ELPEQPQQF LMW glutenin and DQ2.5) 257 omega5-liadin/ Glu-5 minimal epitope in Deamidated form Immunogenic Deamidated DQ2 (DQ2.2 44 9 ELPEQPQQF LMW glutenin and DQ2.5) 258 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQISQPQIPQQQQI or LMW PQQPQQF glutenin 259 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQQQQI or LMW PQQPQQF glutenin 260 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQQQQI or LMW PQQPQQF glutenin 261 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQIPQEQQI or LMW PQQPQQF glutenin 262 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQIPQQQEI or LMW PQQPQQF glutenin 263 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQQQQI or LMW PQQPQQF glutenin 264 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQEQQI or LMW PQQPQQF glutenin 265 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQQQEI or LMW PQQPQQF glutenin 266 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQEQQI or LMW PQQPQQF glutenin 267 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQQQEI or LMW PQQPQQF glutenin 268 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQIPQEQEI or LMW PQQPQQF glutenin 269 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQEQQI or LMW PQQPQQF glutenin 270 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQQQEI or LMW PQQPQQF glutenin 271 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQEQEI or LMW PQQPQQF glutenin 272 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQEQEI or LMW PQQPQQF glutenin 273 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQEQEI or LMW PQQPQQF glutenin 274 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQISQPQLPQQQQI or LMW PQQPQQF glutenin 275 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQQQQI or LMW PQQPQQF glutenin 276 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQQQQI or LMW PQQPQQF glutenin 277 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQLPQEQQI or LMW PQQPQQF glutenin 278 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQLPQQQEI or LMW PQQPQQF glutenin 279 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQQQQI or LMW PQQPQQF glutenin 280 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQEQQI or LMW PQQPQQF glutenin 281 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQQQEI or LMW PQQPQQF glutenin 282 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQEQQI or LMW PQQPQQF glutenin 283 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQQQEI or LMW PQQPQQF glutenin 284 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQLPQEQEI or LMW PQQPQQF glutenin 285 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQEQQI or LMW PQQPQQF glutenin 286 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQQQEI or LMW PQQPQQF glutenin 287 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQEQEI or LMW PQQPQQF glutenin 288 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQEQEI or LMW PQQPQQF glutenin 289 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQEQEI or LMW PQQPQQF glutenin 290 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQISQPQIPQQQQL or LMW PQQPQQF glutenin 291 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQQQQL or LMW PQQPQQF glutenin 292 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQQQQL or LMW PQQPQQF glutenin 293 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQIPQEQQL or LMW PQQPQQF glutenin 294 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQIPQQQEL or LMW PQQPQQF glutenin 295 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQQQQL or LMW PQQPQQF glutenin 296 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQEQQL or LMW PQQPQQF glutenin 297 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQQQEL or LMW PQQPQQF glutenin 298 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQEQQL or LMW PQQPQQF glutenin 299 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQQQEL or LMW PQQPQQF glutenin 300 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQIPQEQEL or LMW PQQPQQF glutenin 301 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQEQQL or LMW PQQPQQF glutenin 302 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQQQEL or LMW PQQPQQF glutenin 303 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQIPQEQEL or LMW PQQPQQF glutenin 304 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPEIPQEQEL or LMW PQQPQQF glutenin 305 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPEIPQEQEL or LMW PQQPQQF glutenin 306 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQLSQPQIPQQQQI or LMW PQQPQQF glutenin 307 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQQQQI or LMW PQQPQQF glutenin 308 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQQQQI or LMW PQQPQQF glutenin 309 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQIPQEQQI or LMW PQQPQQF glutenin 310 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQIPQQQEI or LMW PQQPQQF glutenin 311 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQQQQI or LMW PQQPQQF glutenin 312 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQEQQI or LMW PQQPQQF glutenin 313 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQQQEI or LMW PQQPQQF glutenin 314 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQEQQI or LMW PQQPQQF glutenin 315 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQQQEI or LMW PQQPQQF glutenin 316 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQIPQEQEI or LMW PQQPQQF glutenin 317 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQEQQI or LMW PQQPQQF glutenin 318 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQQQEI or LMW PQQPQQF glutenin 319 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQEQEI or LMW PQQPQQF glutenin 320 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQEQEI or LMW PQQPQQF glutenin 321 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQEQEI or LMW PQQPQQF glutenin 322 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQLSQPQLPQQQQI or LMW PQQPQQF glutenin 323 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQQQQI or LMW PQQPQQF glutenin 324 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQQQQI or LMW PQQPQQF glutenin 325 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQLPQEQQI or LMW PQQPQQF glutenin 326 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQLPQQQEI or LMW PQQPQQF glutenin 327 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQQQQI or LMW PQQPQQF glutenin 328 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQEQQI or LMW PQQPQQF glutenin 329 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQQQEI or LMW PQQPQQF glutenin 330 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQEQQI or LMW PQQPQQF glutenin 331 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQQQEI or LMW PQQPQQF glutenin 332 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQLPQEQEI or LMW PQQPQQF glutenin 333 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQEQQI or LMW PQQPQQF glutenin 334 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQQQEI or LMW PQQPQQF glutenin 335 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQEQEI or LMW PQQPQQF glutenin 336 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQEQEI or LMW PQQPQQF glutenin 337 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQEQEI or LMW PQQPQQF glutenin 338 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQLSQPQIPQQQQL or LMW PQQPQQF glutenin 339 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQQQQL or LMW PQQPQQF glutenin 340 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQQQQL or LMW PQQPQQF glutenin 341 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQIPQEQQL or LMW PQQPQQF glutenin 342 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQIPQQQEL or LMW PQQPQQF glutenin 343 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQQQQL or LMW PQQPQQF glutenin 344 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQEQQL or LMW PQQPQQF glutenin 345 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQQQEL or LMW PQQPQQF glutenin 346 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQEQQL or LMW PQQPQQF glutenin 347 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQQQEL or LMW PQQPQQF glutenin 348 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQIPQEQEL or LMW PQQPQQF glutenin 349 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQEQQL or LMW PQQPQQF glutenin 350 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQQQEL or LMW PQQPQQF glutenin 351 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQIPQEQEL or LMW PQQPQQF glutenin 352 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPEIPQEQEL or LMW PQQPQQF glutenin 353 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPEIPQEQEL or LMW PQQPQQF glutenin 354 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQISQPQLPQQQQL or LMW PQQPQQF glutenin 355 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQQQQL or LMW PQQPQQF glutenin 356 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQQQQL or LMW PQQPQQF glutenin 357 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQLPQEQQL or LMW PQQPQQF glutenin 358 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQLPQQQEL or LMW PQQPQQF glutenin 359 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQQQQL or LMW PQQPQQF glutenin 360 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQEQQL or LMW PQQPQQF glutenin 361 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQQQEL or LMW PQQPQQF glutenin 362 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQEQQL or LMW PQQPQQF glutenin 363 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQQQEL or LMW PQQPQQF glutenin 364 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPQLPQEQEL or LMW PQQPQQF glutenin 365 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQEQQL or LMW PQQPQQF glutenin 366 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQQQEL or LMW PQQPQQF glutenin 367 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPQLPQEQEL or LMW PQQPQQF glutenin 368 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQISQPELPQEQEL or LMW PQQPQQF glutenin 369 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QEISQPELPQEQEL or LMW PQQPQQF glutenin 370 gamma-gliadin Glu-5 Immunogenic Native DQ2 19 21 QQLSQPQLPQQQQL or LMW PQQPQQF glutenin 371 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQQQQL or LMW PQQPQQF glutenin 372 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQQQQL or LMW PQQPQQF glutenin 373 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQLPQEQQL or LMW PQQPQQF glutenin 374 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQLPQQQEL or LMW PQQPQQF glutenin 375 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQQQQL or LMW PQQPQQF glutenin 376 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQEQQL or LMW PQQPQQF glutenin 377 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQQQEL or LMW PQQPQQF glutenin 378 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQEQQL or LMW PQQPQQF glutenin 379 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQQQEL or LMW PQQPQQF glutenin 380 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPQLPQEQEL or LMW PQQPQQF glutenin 381 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQEQQL or LMW PQQPQQF glutenin 382 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQQQEL or LMW PQQPQQF glutenin 383 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPQLPQEQEL or LMW PQQPQQF glutenin 384 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QQLSQPELPQEQEL or LMW PQQPQQF glutenin 385 gamma-gliadin Glu-5 in Deamidated form Immunogenic Deamidated DQ2 19 21 QELSQPELPQEQEL or LMW PQQPQQF glutenin 386 gamma-gliadin gamma-gliadin P08079 Immunogenic Native DQ8 54 20 QQFLQPQQPFPQQP QQPYPQ 387 gamma-gliadin gamma-gliadin P08079 Immunogenic Native DQ8 54 17 QQFLQPQQPFPQQP QQP 388 gamma-gliadin gamma-5 gliadin (p59-p84) Immunogenic Native DQ2 48 26 FLQPQQPFPQQPQQ PYPQQPQQPFPQ 389 gamma-gliadin gamma-5 gliadin (p59-p84; E63) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPQQPQQ PYPQQPQQPFPQ 390 gamma-gliadin gamma-5 gliadin (p59-p84; E68) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPEQPQQ PYPQQPQQPFPQ 391 gamma-gliadin gamma-5 gliadin (p59-p84; E71) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPEQ PYPQQPQQPFPQ 392 gamma-gliadin gamma-5 gliadin (p59-p84; E76) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPQQ PYPEQPQQPFPQ 393 gamma-gliadin gamma-5 gliadin (p59-p84; E79) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPQQ PYPQQPEQPFPQ 394 gamma-gliadin gamma-5 gliadin (p59-p84; E63 and E68) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPEQPQQ PYPQQPQQPFPQ 395 gamma-gliadin gamma-5 gliadin (p59-p84; E63 and E71) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPQQPEQ PYPQQPQQPFPQ 396 gamma-gliadin gamma-5 gliadin (p59-p84; E63 and E76) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPQQPQQ PYPEQPQQPFPQ 397 gamma-gliadin gamma-5 gliadin (p59-p84; E63 and E79) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPQQPQQ PYPQQPEQPFPQ 398 gamma-gliadin gamma-5 gliadin (p59-p84; E68 and E71) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPEQPEQ PYPQQPQQPFPQ 399 gamma-gliadin gamma-5 gliadin (p59-p84; E68 and E76) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPEQPQQ PYPEQPQQPFPQ 400 gamma-gliadin gamma-5 gliadin (p59-p84; E68 and E79) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPEQPQQ PYPQQPEQPFPQ 401 gamma-gliadin gamma-5 gliadin (p59-p84; E71 and E76) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPEQ PYPEQPQQPFPQ 402 gamma-gliadin gamma-5 gliadin (p59-p84; E71 and E79) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPEQ PYPQQPEQPFPQ 403 gamma-gliadin gamma-5 gliadin (p59-p84; E76 and E79) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPQQ PYPEQPEQPFPQ 404 gamma-gliadin gamma-5 gliadin (p59-p84; E63, E68 and E71) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPEQPEQ PYPQQPQQPFPQ 405 gamma-gliadin gamma-5 gliadin (p59-p84; E63, E68 and E76) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPEQPQQ PYPEQPQQPFPQ 406 gamma-gliadin gamma-5 gliadin (p59-p84; E63, E68 and E79) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPEQPQQ PYPQQPEQPFPQ 407 gamma-gliadin gamma-5 gliadin (p59-p84; E68, E71 and E76) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPEQPEQ PYPEQPQQPFPQ 408 gamma-gliadin gamma-5 gliadin (p59-p84; E68, E71 and E79) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPEQPEQ PYPQQPEQPFPQ 409 gamma-gliadin gamma-5 gliadin (p59-p84; E71, E76 and E79) Immunogenic Deamidated DQ2 48 26 FLQPQQPFPQQPEQ PYPEQPEQPFPQ 410 gamma-gliadin gamma-5 gliadin (p59-p84; E63, E68, E71, E76 and E79) Immunogenic Deamidated DQ2 48 26 FLQPEQPFPEQPEQ PYPEQPEQPFPQ 411 gamma-gliadin  gamma-5 gliadin (p60-p79); DQ2-gamma-V gamma-Glia Immunogenic Native DQ2 57, 23, 48, 20 LQPQQPFPQQPQQP (p78-p97); gamma-3 and gamma-5 peptide 1317 2 YPQQPQ 412 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPQQPQQP YPQQPQ 413 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E86) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPEQPQQP YPQQPQ 414 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E89) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPQQPEQP YPQQPQ 415 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E94) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPQQPQQP YPEQPQ 416 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81 and E86) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPEQPQQP YPQQPQ 417 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81 and E89) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPQQPEQP YPQQPQ 418 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81 and E94) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPQQPQQP YPEQPQ 419 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E86 and E89) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPEQPEQP YPQQPQ 420 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E86 and E64) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPEQPQQP YPEQPQ 421 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E89 and E94) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPQQPEQP YPEQPQ 422 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81, E86 and E89) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPEQPEQP YPQQPQ 423 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81, E86 and E94) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPEQPQQP YPEQPQ 424 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E86, E89 and E94) Immunogenic Deamidated DQ2 57, 23, 2 20 LQPQQPFPEQPEQP YPEQPQ 425 gamma-gliadin DQ2-gamma-V gamma-Glia (p78-p97; E81, E86, E89 and Immunogenic Deamidated DQ2 57, 23, 2 20 LQPEQPFPEQPEQP E94) YPEQPQ 426 gamma-gliadin gamma3/gamma4 Immunogenic Native DQ2 25 17 PQQPFPQQPQQPYP QQP 427 gamma-gliadin gamma5 (p62-p74) Immunogenic Native DQ2 25 13 PQQPFPQQPQQPY 428 gamma-gliadin gamma5 (p62-p74; E68) Immunogenic Deamidated DQ2 25 13 PQQPFPEQPQQPY 429 gamma-gliadin gamma5 (p62-p74; E63 and E68) Immunogenic Deamidated DQ2 25 13 PEQPFPEQPQQPY 430 gamma-gliadin gamma5 (p62-p74; E68 and E71) Immunogenic Deamidated DQ2 25 13 PQQPFPEQPEQPY 431 gamma-gliadin gamma5 (p62-p74; E63, E68 and E71) Immunogenic Deamidated DQ2 25 13 PEQPFPEQPEQPY 432 gamma-gliadin gamma-5 gliadin (p62-p72) Immunogenic Native DQ2 47, 48 11 PQQPFPQQPQQ 433 gamma-gliadin gamma5-gliadin (p62-p72; E68) Immunogenic Deamidated DQ2 47 11 PQQPFPEQPQQ 434 gamma-gliadin gamma5 (p62-p72; E68, E63 and E71) Immunogenic Deamidated DQ2 25 11 PEQPFPEQPEQ 435 gamma-gliadin gamma23mer Immunogenic Native DQ2.5 61 23 QQPFPQQPQQPYPQ QPQQPFPQP 436 gamma-gliadin gamma23mer (in considered Deamidated form) Immunogenic Deamidated DQ2.5 61 23 EQPFPEQPEQPYPE QPEQPFPQP 437 gamma-gliadin gamma5-gliadin (p60-p79) Immunogenic Native DQ2 21 14 QQPFPQQPQQPYPQ 438 gamma-5 gliadin DQ2.5_glia_gamma5 Immunogenic Native DQ2.5 25, 90, 23 9 QQPFPQQPQ 439 gamma-5 gliadin DQ2.5_glia_gamma5 Immunogenic Deamidated DQ2.5 25, 90, 23 9 QQPFPEQPQ 440 gamma-gliadin CAUTION 100% match to Archaea protein lower to others Immunogenic Deamidated DQ2 25 9 EQPFPEQPE and to gamma5 (p63-p71; E63, E68 and E71) 441 gamma-gliadin DQ2-gamma-Ill gamma-Glia (p83-p97) Immunogenic Native DQ2 57, 48, 23, 15 PFPQQPQQPYPQQP Q 2 442 gamma-gliadin DQ2-gamma-Ill gamma-Glia (p83-p97; E86) Immunogenic Deamidated DQ2 57, 48, 23, 15 PFPEQPQQPYPQQP Q 2 443 gamma-gliadin DQ2-gamma-Ill gamma-Glia (p83-p97; E89) Immunogenic Deamidated DQ2 57, 48, 23, 15 PFPQQPEQPYPQQP Q 2 444 gamma-gliadin DQ2-gamma-Ill gamma-Glia (p83-p97; E86 and E89) Immunogenic Deamidated DQ2 57, 48, 23, 15 PFPEQPEQPYPQQP Q 2 445 gamma-gliadin Wheat peptide W23 Immunogenic Native DQ2 62 12 PFPQQPQQPYPQ 446 gamma-gliadin gamma-gliadin (p66-p80) AJ416339 Immunogenic Native DQ8, DQ2 17 15 FPQQPQQPYPQQPQ Q 447 gamma-gliadin gamma-gliadin (p66-p80; E68) AJ416339 Immunogenic Deamidated DQ8 17, 83 15 FPEQPQQPYPQQPQ Q 448 gamma-gliadin gamma-gliadin (p66-p80; E71) AJ416339 Immunogenic Deamidated DQ2 17 15 FPQQPEQPYPQQPQ Q 449 gamma-gliadin gamma-gliadin (p66-p80; E76) AJ416339 Immunogenic Deamidated DQ8, DQ2 17 15 FPQQPQQPYPEQPQ Q 450 gamma-gliadin  gamma-gliadin (p66-p80; E71 and 76) AJ416339 Immunogenic Deamidated DQ8 17 15 FPEQPQQPYPEQPQ Q 451 gamma-gliadin gamma5 (p66-p78) Immunogenic Native DQ2 23 13 FPQQPQQPYPQQP 452 gamma-gliadin  gamma5 (p66-p78; E68 and E71) Immunogenic Deamidated DQ2 23 13 FPEQPEQPYPQQP 453 gamma-gliadin gamma5 (p66-p78; E68 and E72) Immunogenic Deamidated DQ2 23 13 FPEQPQEPYPQQP 454 gamma-gliadin gamma5 (p66-p77) Immunogenic Native DQ2 25 12 FPQQPQQPYPQQ 455 gamma-gliadin gamma5 (p66-p77; E71) Immunogenic Deamidated DQ2 25 12 FPQQPEQPYPQQ 456 gamma-gliadin  gamma5 (p66-p77; E68, E71 and E76) Immunogenic Deamidated DQ2 25 12 FPEQPEQPYPEQ 457 gamma-gliadin  gamma5 (p67-p77; E68, E71 and E76) Immunogenic Deamidated DQ2 25 11 PEQPEQPYPEQ 458 gamma-1 and CAUTION 100% matches to many fungal and parasite Immunogenic Native DQ2.5/DQ8 17, 23, 78 9 QQPQQPYPQ gamma5 gliadin proteins as well as DQ2.5_glia_gamma3 DQ8_glia_gamma 1b 459 gamma-5 gliadin DQ8_glia_gamma1b DQ2.5_glia_gamma3 Immunogenic Deamidated DQ2.5/DQ8 25, 17, 78 9 EQPEQPYPE 460 gamma-1 gliadin DQ8_glia_gamma 1b Immunogenic Deamidated DQ2.5/DQ8 25, 17, 78 9 EQPQQPYPE 461 gamma-1 gliadin DQ8_glia_gamma 1b Immunogenic Deamidated DQ2.5/DQ8 17, 78 9 EQPQQPFPE 462 gamma-III CAUTION 100% identity matches to fungal and lower to Immunogenic Deamidated DQ2.5/DQ8 17, 78 9 QQPEQPYPQ gliadin other fungal proteins as DQ2.5_glia_gamma3 DQ8_glia_gamma1b 463 gamma-gliadin Predicted gamma-gliadin peptide Immunogenic Native DQ8 (DQ2/8) 22 14 QQPYPQQPQQPFPQ 464 gamma-gliadin CAUTION many 100% identity matches to legume and Immunogenic Native DQ2 25 9 QQPYPQQPQ   bacterial proteins and also gamma-Vib gliadin 465 gamma-gliadin CAUTION 100% match to bacterial protein and lower to Immunogenic Deamidated DQ2 25 9 EQPYPQQPQ others and gamma-Vib gliadin in Deamidated form 466 gamma-gliadin CAUTION 100% identity to bacterial protein and lower Immunogenic Deamidated DQ2 25 9 QQPYPEQPQ to others and gamma-Vib gliadin in Deamidated form 467 gamma-gliadin gamma-Vib gliadin in Deamidated form Immunogenic Deamidated DQ2 25 9 EQPYPEQPQ 468 gamma-gliadin CAUTION many 100% matches to fish, fungi and other Immunogenic Native DQ2 (DQ2.2 44 9 PYPQQPQQP non-wheat family proteins also a few to Secalin and DQ2.5) Glia-gamma2 469 gamma-gliadin Glia-gamma2 in Deamidated form Immunogenic Deamidated DQ2 (DQ2.2 44, 83 9 PYPEQPQQP and DQ2.5) 470 gamma-gliadin Glia-gamma2 in Deamidated form Immunogenic Deamidated DQ2 (DQ2.2 44 9 PYPQQPEQP and DQ2.5) 471 gamma-gliadin CAUTION 100% match to bacterial protein and lizard Immunogenic Deamidated DQ2 (DQ2.2 44 9 PYPEQPEQP protein lower to grapes and wheat Glia-gamma2 in and DQ2.5) Deamidated form 472 gamma-gliadin CAUTION 100% 4 matches to Candida 2 to bacteria and Immunogenic Native DQ2.5/DQ8 90, 17, 8 9 QQPQQPFPQ many to secalins and wheat gamma2-gliadin  473 gamma-2 gliadin CAUTION 100% match to fungal protein and lower to Immunogenic Deamidated DQ2.5/DQ8 90, 17, 8 9 EQPQQPFPQ Candida and to Gliadin epitope: gamma-1 474 gamma-2 gliadin Gliadin epitope: gamma-VII Immunogenic Deamidated DQ2.5/DQ8 90, 17, 8 9 QQPEQPFPQ 475 gamma-2 gliadin CAUTION 100% match to a bacterial protein/lower to Immunogenic Deamidated DQ2.5/DQ8 25, 90, 17 9 EQPEQPFPQ   triticum gammaVII-gliadin in Deamidated form 476 gamma-gliadin gamma-Glia (p105-p118) Immunogenic Native DQ2 57 14 PQQQTLQPQQPAQL 477 gamma-gliadin gamma-Glia (p105-p118; E113) Immunogenic Deamidated DQ2 57 14 PQQQTLQPEQPAQL 478 gamma1-gliadin Wheat peptide W37 Immunogenic Native DQ2 62 20 ATANMQVDPSGQVQ WPQQQP 479 gamma1-gliadin Wheat peptide W37 Immunogenic Native DQ2 62 12 QVDPSGQVQWPQ 480 gamma1-gliadin gamma-gliadin 1370 (p1-p30); gamma-gliadin M2 M36999 Immunogenic Native DQ2, DQ8 17, 23 20 WPQQQPFPQPQQPF (p11-p30) homologous to DQ2-alpha-I CQQPQR 481 gamma1-gliadin gamma-gliadin 1371 (p21-p40) Immunogenic Native DQ2 17 20 QQPFCQQPQRTIPQ PHQTFH 482 gamma1-gliadin gamma-gliadin 1372 (p31-p50) Immunogenic Native DQ2 17 20 TIPQPHQTFHHQPQ QTFPQP 483 gamma1-gliadin gamma-gliadin 1372 (p41-p60) Immunogenic Native DQ2, DQ8 17 20 HQPQQTFPQPQQTY PHQPQQ 484 gamma1-gliadin gamma-gliadin 1372 (p51-p70) Immunogenic Native DQ2 17 20 QQTYPHQPQQQFPQ TQQPQQ 485 gamma1-gliadin gamma-gliadin 1375 (p61-p80); gamma-gliadin M7 M36999 Immunogenic Native DQ2, DQ8 25, 17, 23 20 QFPQTQQPQQPFPQ (p61-p80) homologous to DQ2-gamma-III PQQTFP 486 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64); gamma-gliadin M7 Immunogenic Deamidated DQ2 25 20 QFPETQQPQQPFPQ M36999 (p61-p80) homologous to DQ2-gamma-III PQQTFP 487 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E66); gamma-gliadin M7 Immunogenic Deamidated DQ2 25 20 QFPQTEQPQQPFPQ M36999 (p61-p80) homologous to DQ2-gamma-III PQQTFP 488 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E69); gamma-gliadin M7 Immunogenic Deamidated DQ2 25 20 QFPQTQQPEQPFPQ M36999 (p61-p80) homologous to DQ2-gamma-III PQQTFP 489 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E76); gamma-gliadin M7 Immunogenic Deamidated DQ2 25 20 QFPQTQQPQQPFPQ M36999 (p61-p80) homologous to DQ2-gamma-III PEQTFP 490 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64 and E66); gamma- Immunogenic Deamidated DQ2 25 20 QFPETEQPQQPFPQ gliadin M7 M36999 (p61-p80) homologous to PQQTFP DQ2-gamma-III 491 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64 and E69); gamma- Immunogenic Deamidated DQ2 25 20 QFPETQQPEQPFPQ gliadin M7 M36999 (p61-p80) homologous to PQQTFP DQ2-gamma-III 492 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64 and E76); gamma- Immunogenic Deamidated DQ2 25 20 QFPETQQPQQPFPQ gliadin M7 M36999 (p61-p80) homologous to PEQTFP DQ2-gamma-III 493 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E66 and E69); gamma- Immunogenic Deamidated DQ2 25 20 QFPQTEQPEQPFPQ gliadin M7 M36999 (p61-p80) homologous to PQQTFP DQ2-gamma-III 494 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E66 and E76); gamma- Immunogenic Deamidated DQ2 25 20 QFPQTEQPQQPFPQ gliadin M7 M36999 (p61-p80) homologous to PEQTFP DQ2-gamma-III 495 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E69 and E76); gamma- Immunogenic Deamidated DQ2 25 20 QFPQTQQPEQPFPQ gliadin M7 M36999 (p61-p80) homologous to PEQTFP DQ2-gamma-III 496 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64, E66 and E69); Immunogenic Deamidated DQ2 25 20 QFPETEQPEQPFPQ gamma-gliadin M7 M36999 (p61-p80) homologous to PQQTFP DQ2-gamma-III 497 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64, E66 and E76); Immunogenic Deamidated DQ2 25 20 QFPETEQPQQPFPQ gamma-gliadin M7 M36999 (p61-p80) homologous to PEQTFP DQ2-gamma-III 498 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64, E69 and E76); Immunogenic Deamidated DQ2 25 20 QFPETQQPEQPFPQ gamma-gliadin M7 M36999 (p61-p80) homologous to PEQTFP DQ2-gamma-III 499 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E66, E69 and E76); Immunogenic Deamidated DQ2 25 20 QFPQTEQPEQPFPQ gamma-gliadin M7 M36999 (p61-p80) homologous to PEQTFP DQ2-gamma-III 500 gamma1-gliadin gamma-gliadin 1375 (p61-p80; E64, E66, E69 and Immunogenic Deamidated DQ2 25 20 QFPETEQPEQPFPQ E76); gamma-gliadin M7 M36999 (p61-p80) PEQTFP homologous to DQ2-gamma-III 501 gamma1-gliadin Wheat peptide W28, W33 Immunogenic Native DQ2 62 20 PQQPFPQPQQTFPQ QPQLPF 502 gamma1-gliadin gamma-gliadin 1376 (p71-p90); gamma-gliadin M8 Immunogenic Native DQ2, DQ8 17, 23 20 PFPQPQQTFPQQPQ M36999 (71-80) homologous to DQ2-alpha-I and LPFPQQ DQ2-gamma-IV 503 gamma1-gliadin Wheat peptide W33 Immunogenic Native DQ2 62 11 PFPQPQQTFPQ 504 gamma1-gliadin Wheat peptide W28 Immunogenic Native DQ2 62 12 PQQTFPQQPQLP 505 gamma1-gliadin Wheat peptide W10 Immunogenic Native DQ2 62 20 SQQPQQQFSQPQQQ FPQPQQ 506 gamma1-gliadin DQ2-y-IV y-Glia (p117-p132) Immunogenic Native DQ2 57, 23 15 QQFSQPQQQFPQPQ Q 507 gamma1-gliadin DQ2-y-IV y-Glia (p117-p132; E123) Immunogenic Deamidated DQ2 57, 23 15 QQFSQPEQQFPQPQ Q 508 gamma1-gliadin DQ2-y-IV y-Glia (p117-p132; E125) Immunogenic Deamidated DQ2 57, 23 15 QQFSQPQQEFPQPQ Q 509 gamma1-gliadin DQ2-y-IV y-Glia (p117-p132; E123 and E125) Immunogenic Deamidated DQ2 57, 23 15 QQFSQPEQEFPQPQ Q 510 gamma1-gliadin Wheat peptide W10 Immunogenic Native DQ2 62 12 QQFSQPQQQFPQ 511 gamma1-gliadin gamma-IV (p101-p113) Immunogenic Native DQ2 23 13 QFSQPQQQFPQPQ 512 gamma1-gliadin gamma-gliadin (gammas p102-p113) Immunogenic Native DQ2 23 12 FSQPQQQFPQPQ 513 gamma1-gliadin gamma5 (p102-p113; E106) Immunogenic Deamidated DQ2 23 12 FSQPEQQFPQPQ 514 gamma1-gliadin gamma5 (p102-p113; E108) Immunogenic Deamidated DQ2 23 12 FSQPQQEFPQPQ 515 gamma1-gliadin gamma5 (p102-p113; E106 and E108) Immunogenic Deamidated DQ2 25, 23 12 FSQPEQEFPQPQ 516 gamma1-gliadin gamma5 (p102-p111) Immunogenic Native DQ2 25 10 FSQPQQQFPQ 517 gamma1-gliadin gamma5 (p102-p111; E106) Immunogenic Deamidated DQ2 25 10 FSQPEQQFPQ 518 gamma1-gliadin gamma5 (p102-p111; E108) Immunogenic Deamidated DQ2 25 10 FSQPQQEFPQ 519 gamma1-gliadin gamma5 (p102-p111; E106 and E108) Immunogenic Deamidated DQ2 25 10 FSQPEQEFPQ 520 gamma1-gliadin gamma-IV (p103-p114) Immunogenic Native DQ2 23 12 SQPQQQFPQPQQ 521 gamma-5 gliadin CAUTION 100% match to fungal protein and many Immunogenic Native DQ2.5 90, 25, 23 9 SQPQQQFPQ wheat proteins glia-gamma 4a 522 gamma-5 gliadin glia-gamma 4a Immunogenic Deamidated DQ2.5 90, 25, 23 9 SQPEQEFPQ 523 gamma1-gliadin gamma-gliadin of GDB2_WHEAT (SwissProt P08453 Immunogenic Native DQ2 24 11 QQPQQSFPQQQ GI:121101) (p140-p150) 524 gamma1-gliadin gamma-gliadin of GDB2_WHEAT (SwissProt P08453 Immunogenic Native DQ2 24 10 QPQQSFPQQQ GI170738) (p141-p150) 525 gamma1-gliadin gamma-gliadin of GDB2_WHEAT (SwissProt P08453 Immunogenic Deamidated DQ2 24 10 QPQQSFPEQQ GI170738) (p141-p150; E148) 526 gamma-gliadin Wheat peptide W07 Immunogenic Native DQ2 62 20 WPQQQPFPQPQQPF CQQPQQ 527 gamma-gliadin Wheat peptide W07 Immunogenic Deamidated DQ2 62 20 WPQQQPFPQPEQPF CQQPQQ 529 gamma-gliadin Wheat peptide W07 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPFCQ 530 gamma-gliadin Predicted gamma-gliadin Immunogenic Native DQ8 (DQ2/8) 22 14 QFPQTQQPQQPFPQ 531 gamma-gliadin gamma-gliadin M36999 (p63-p76; E66) Immunogenic Synthesised DQ8 17 14 PQTEQPQQPFPQPQ as Deamidated 532 gamma-gliadin gamma-gliadin M36999 (p63-p76; E69) Immunogenic Synthesised DQ2 17 14 PQTQQPEQPFPQPQ as Deamidated 533 gamma-gliadin gamma-gliadin M36999 (p63-p76; E66 and E69) Immunogenic Synthesised DQ2, DQ8 17 14 PQTEQPEQPFPQPQ as Deamidated 534 gamma-gliadin gamma-gliadin 1375 (p61-p80); gamma-gliadin M7 Immunogenic Native DQ2 25 11 TQQPQQPFPQP M36999 (61-80) homologous to DQ2-gamma-III 535 gamma-gliadin gamma-gliadin 1375 (p61-p80; E62 and E65),; gamma- Immunogenic Deamidated DQ2 25 11 TEQPEQPFPQP gliadin M7 M36999 (61-80) homologous to DQ2-gamma-III 536 gamma-gliadin gamma-gliadin 1377 (p81-p100) Immunogenic Native DQ2, DQ8 17 20 QQPQLPFPQQPQQP FPQPQQ 537 gamma-gliadin gamma-gliadin (p84-p97) Immunogenic Native DQ8 (DQ2/8) 22 14 QLPFPQQPQQPFPQ 538 gamma-gliadin Glia-gamma2 (p89-p102) Immunogenic Native DQ2 20 10 PFPQQPQQPF 539 gamma-gliadin Glia-gamma2 (p89-p102; E92) Immunogenic Deamidated DQ2 20 10 PFPEQPQQPF 540 gamma-gliadin Glia-gamma2 (p89-p102; E94) Immunogenic Deamidated DQ2 20 10 PFPQQPEQPF 541 gamma-gliadin Glia-gamma2 (p89-p102; E92 and E94) Immunogenic Deamidated DQ2 20 10 PFPEQPEQPF 542 gamma-gliadin CAUTION 100% match to a fungal protein and many Immunogenic Native DQ2 27 9 FPQQPQQPF Secalins gamma-Gliadin (p90-p102) 543 gamma-gliadin gamma-Gliadin (p90-p102; E92) Immunogenic Deamidated DQ2 27 9 FPEQPQQPF 544 gamma-gliadin gamma-Gliadin (p90-p102; E96) Immunogenic Deamidated DQ2 27 9 FPQQPQEPF 545 gamma-gliadin gamma-Gliadin (p90-p102; E92 and E96) Immunogenic Deamidated DQ2 27 9 FPEQPQEPF 546 gamma-gliadin gamma-gliadin 1378 (p91-p110), ; gamma-gliadin Immunogenic Native DQ2, DQ8 17, 23 20 PQQPFPQPQQPQQP M10 M36999 (91-110) homologous to DQ2-alpha-I FPQSQQ 547 gamma-gliadin Wheat peptide W36 Immunogenic Native DQ2 62 20 QQPAQYEVIRSLVL RTLPNM 548 gamma-gliadin Wheat peptide W36 Immunogenic Native DQ2 62 16 QYEVIRSLVLRTLP NM 549 gamma-gliadin Wheat peptide W36 Immunogenic Deamidated DQ2 62 15 EYEVIRSLVLRTLP N 550 gamma-gliadin Wheat peptide W36 Immunogenic Native DQ2 62 15 QYQVIRSLVLRTLP N 551 gamma-gliadin gamma-gliadin AAK84778 (p74-p93) Immunogenic Native DQ8 54 20 QQQFIQPQQPFPQQ PQQTYP 552 gamma-gliadin Wheat peptide W14 Immunogenic Native DQ2 62 12 QQFIQPQQPFPQ 553 gamma-gliadin Predicted gamma-gliadin Immunogenic Native DQ8 (DQ2/8) 22 14 PFPQTQQPQQPFPQ 554 gamma-gliadin gamma-gliadin 1379 (p101-p120) Immunogenic Native DQ2, DQ8 17 20 PQQPFPQSQQPQQP FPQPQQ 555 gamma-gliadin Predicted gamma-gliadin Immunogenic Native DQ8 (DQ2/8) 22 14 PFPQSQQPQQPFPQ 556 gamma-gliadin Wheat peptide W16 Immunogenic Native DQ2 62 20 SQQPQQPFPQPQQQ FPQPQQ 557 gamma-gliadin gamma-gliadin 1380 (p111-p130); gamma-gliadin M12 Immunogenic Native DQ2 25, 17, 23 20 PQQPFPQPQQQFPQ M36999 (111-130) homologous to DQ2-gamma-IV PQQPQQ 558 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112); gamma-gliadin Immunogenic Deamidated DQ2 25 20 PEQPFPQPQQQFPQ M12 M36999 (111-130) homologous to DQ2-gamma-IV PQQPQQ 559 gamma-gliadin gamma-gliadin 1380 (p111-p130; E119); gamma-gliadin Immunogenic Deamidated DQ2 25 20 PQQPFPQPEQQFPQ M12 M36999 (111-130) homologous to DQ2-gamma-IV PQQPQQ 560 gamma-gliadin gamma-gliadin 1380 (p111-p130; E121); gamma-gliadin Immunogenic Deamidated DQ2 25 20 PQQPFPQPQQEFPQ M12 M36999 (111-130) homologous to DQ2-gamma-IV PQQPQQ 561 gamma-gliadin gamma-gliadin 1380 (p111-p130; E126); gamma-gliadin Immunogenic Deamidated DQ2 25 20 PQQPFPQPQQQFPQ M12 M36999 (111-130) homologous to DQ2-gamma-IV PEQPQQ 562 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112 and E119); gamma- Immunogenic Deamidated DQ2 25 20 PEQPFPQPEQQFPQ gliadin M12 M36999 (111-130) homologous to PQQPQQ DQ2-gamma-IV 563 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112 and E121); gamma- Immunogenic Deamidated DQ2 25 20 PEQPFPQPQQEFPQ gliadin M12 M36999 (111-130) homologous to PQQPQQ DQ2-gamma-IV 564 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112 and E126); gamma- Immunogenic Deamidated DQ2 25 20 PEQPFPQPQQQFPQ gliadin M12 M36999 (111-130) homologous to PEQPQQ DQ2-gamma-IV 565 gamma-gliadin gamma-gliadin 1380 (p111-p130; E119 and E121); gamma- Immunogenic Deamidated DQ2 25 20 PQQPFPQPEQEFPQ gliadin M12 M36999 (111-130) homologous to PQQPQQ DQ2-gamma-IV 566 gamma-gliadin gamma-gliadin 1380 (p111-p130; E119 and E126); gamma- Immunogenic Deamidated DQ2 25 20 PQQPFPQPEQQFPQ gliadin M12 M36999 (111-130) homologous to PEQPQQ DQ2-gamma-IV 567 gamma-gliadin gamma-gliadin 1380 (p111-p130; E121 and E126); gamma- Immunogenic Deamidated DQ2 25 20 PQQPFPQPQQEFPQ gliadin M12 M36999 (111-130) homologous to PEQPQQ DQ2-gamma-IV 568 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112, E119 and E121); Immunogenic Deamidated DQ2 25 20 PEQPFPQPEQEFPQ gamma- gliadin M12 M36999 (111-130) homologous to PQQPQQ DQ2-gamma-IV 569 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112, E119 and E126); Immunogenic Deamidated DQ2 25 20 PEQPFPQPEQQFPQ gamma-gliadin M12 M36999 (111-130) homologous to PEQPQQ DQ2-gamma-IV 570 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112, E121 and E126); Immunogenic Deamidated DQ2 25 20 PEQPFPQPQQEFPQ gamma-gliadin M12 M36999 (111-130) homologous to PEQPQQ DQ2-gamma-IV 571 gamma-gliadin gamma-gliadin 1380 (p111-p130; E119, E121 and E126); Immunogenic Deamidated DQ2 25 20 PQQPFPQPEQEFPQ gamma-gliadin M12 M36999 (111-130) homologous to PEQPQQ DQ2-gamma-IV 572 gamma-gliadin gamma-gliadin 1380 (p111-p130; E112, E119, E121 and Immunogenic Deamidated DQ2 25 20 PEQPFPQPEQEFPQ E126); gamma-gliadin M12 M36999 (111-130) homologous PEQPQQ to DQ2-gamma-IV 573 gamma-gliadin gamma-gliadin 1380 (p111-p130); gamma-gliadin M12 Immunogenic Native DQ2 62, 25 12 FPQPQQQFPQPQ M36999 (111-130) homologous to DQ2-gamma-IV; W16 574 gamma-gliadin gamma-gliadin 1380 (p111-p130; E115); gamma-gliadin Immunogenic Deamidated DQ2 25 12 FPQPEQQFPQPQ M12 M36999 (111-130) homologous to DQ2-gamma-IV 575 gamma-gliadin gamma-gliadin 1380 (p111-p130; E117); gamma-gliadin Immunogenic Deamidated DQ2 25 12 FPQPQQEFPQPQ M12 M36999 (111-130) homologous to DQ2-gamma-IV 576 gamma-gliadin gamma-gliadin 1380 (p111-p130; E115 and E117); gamma- Immunogenic Deamidated DQ2 25 12 FPQPEQEFPQPQ gliadin M12 M36999 (111-130) homologous to DQ2-gamma-IV 577 gamma-gliadin CAUTION 100% matches to 3 fungal and metazoan Immunogenic Native DQ2.5 90, 25 9 PQPQQQFPQ proteins and wheat glia-gamma 4b 578 gamma-gliadin glia-gamma 4b Immunogenic Deamidated DQ2.5 90, 25 9 PQPEQQFPQ 579 gamma-gliadin glia gamma 4b Immunogenic Deamidated DQ2.5 90, 25 9 PQPQQEFPQ 580 gamma-gliadin glia-gamma 4b Immunogenic Deamidated DQ2.5 90, 25 9 PQPEQEFPQ 581 gamma-gliadin Wheat peptide W16 Immunogenic Deamidated DQ2 62 16 GQQPFPQPEQEFPQ PG 582 gamma-gliadin Wheat peptide W16 Immunogenic Deamidated DQ2 62 13 QPFPQPEQEFPQP 583 gamma-1 glia-alpha 1, glia-gamma 1 Immunogenic Native DQ2.5/DQ8 8, 74, 76, 9 PQQSFPQQQ gliadin 24 584 gamma-gliadin glia-alpha1, glia-gamma1 Immunogenic Deamidated DQ2.5/DQ8 8, 74, 76, 9 PQQSFPQQE 24 585 gamma-1 glia-alpha 1, glia-gamma 1 Immunogenic Deamidated DQ2.5/DQ8 8, 76, 24 9 PQQSFPEQE gliadin 586 gamma-1 glia-alpha1, glia-gamma1 Immunogenic Deamidated DQ2.5, DQ8 8, 17, 76, 9 PQQSFPEQQ gliadin 24 587 gamma-gliadin Glia-gamma30-gliadin (p222-p236) Immunogenic Native DQ2 19 15 VQGQGIIQPQQPAQ L 588 gamma-gliadin Glia-gamma30-gliadin (p222-236; E225) Immunogenic Deamidated DQ2 19 15 VQGEGIIQPQQPAQ L 589 gamma-gliadin Glia-gamma30-gliadin (p222-236; E231) Immunogenic Deamidated DQ2 19 15 VQGQGIIQPEQPAQ L 590 gamma-gliadin Glia-gamma30-gliadin (p222-236; E225 and E231) Immunogenic Deamidated DQ2 19 15 VQGEGIIQPEQPAQ L 591 gamma-gliadin DQ2-y -II y-Glia (p222-p236) Immunogenic Native DQ2 57 15 GQGIIQPQQPAQLI R 592 gamma-gliadin DQ2-y -II y-Glia (p222-p236; E229) Immunogenic Deamidated DQ2 57 15 GQGIIQPEQPAQLI R 593 gamma-gliadin gamma5-gliadin (p227-p237); gamma-ll epitope Immunogenic Native DQ2 25 11 GIIQPQQPAQL 594 gamma-gliadin gamma5-gliadin (p227-237; E232) Immunogenic Deamidated DQ2 25 11 GIIQPEQPAQL 595 gamma-gliadin gamma5-gliadin (p228-237) Immunogenic Native DQ2 47, 25 10 IIQPQQPAQL 596 gamma-gliadin gamma5-gliadin (p228-237; E232) Immunogenic Deamidated DQ2 47, 25 10 IIQPEQPAQL 597 gamma-gliadin gamma-2 peptide 1306; Glia-gamma30-gliadin (p227-p235) Immunogenic Native DQ2 19, 23, 2 9 IIQPQQPAQ minimal epitope 598 gamma-gliadin gamma-2 peptide 1306; Glia-gamma30-gliadin (p228-p235; Immunogenic Deamidated DQ2 19 9 IIQPEQPAQ E232) minimal epitope 599 gamma-5 glia-gamma 2 Immunogenic Native DQ2.5 90, 8, 25 9 IQPQQPAQL gliadin 600 gamma-5 glia-gamma 2 Immunogenic Deamidated DQ2.5 90, 8, 25 9 IQPEQPAQL gliadin 601 gamma-gliadin gamma-gliadin 1381 (p121-p140); gamma-gliadin M13 Immunogenic Native DQ2, DQ8 17, 23 20 QFPQPQQPQQSFPQ M36999 (121-140) identical to DQ2-gamma-I QQQPAI 602 gamma-gliadin DQ2-gamma-I gamma-Glia (p139-p153) Immunogenic Native DQ2 57 15 PQQPQQSFPQQQQP A 603 gamma-gliadin DQ2-gamma-I gamma-Glia (p139-p153; E147) Immunogenic Deamidated DQ2 57 15 PQQPQQSFPEQQQP A 604 gamma-gliadin DQ2-gamma-I gamma-Glia (p139-p153; E150) Immunogenic Deamidated DQ2 57 15 PQQPQQSFPQQEQP A 605 gamma-gliadin DQ2-gamma-I gamma-Glia (p139-p153; E147 and E150) Immunogenic Deamidated DQ2 57 15 PQQPQQSFPEQEQP A 606 gamma-gliadin gamma-gliadin 1382 (p131-p150) Immunogenic Native DQ8 17 20 SFPQQQQPAIQSFL QQQM 607 gamma-gliadin gamma-gliadin 1383 (p141-p160) Immunogenic Native DQ8 17 20 QSFLQQQMNPCKNF LLQQCN 608 gamma-gliadin gamma-gliadin 1388 (p201-p220) Immunogenic Native DQ2 17 20 IHSVAHSIIMQQEQ QQGVPI 609 gamma-gliadin gamma-gliadin M23 M36999 (221-240) homologous to DQ2- Immunogenic Native DQ2 17, 23 20 LRPLFQLAQGLGII gamma-II QPQQPA 610 gamma-gliadin gamma-gliadin 1391 (p231-p250); gamma-gliadin M24 Immunogenic Native DQ2 17, 23 20 LGIIQPQQPAQLEG M36999 (231-250) identical to DQ2-gamma-II IRSLVL 611 gamma-gliadin Gluten peptide E19019 Immunogenic Native DQ2 (DQ2.5) 61 18 PHQPQQQVPQPQQP QQPF 612 gamma-gliadin Predicted gamma-gliadin Immunogenic Native DQ8 (DQ2/8) 22, 8 14 QQPFPQQPQQPFPQ 613 gamma-gliadin Glia-gamma2 in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFPEQPEQPFPQ 614 gamma-gliadin Glia-gamma2 in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFPQQPEQPFPQ 615 gamma-gliadin Glia-gamma2 in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFPEQPQQPFPQ 616 gamma-gliadin gamma-gliadin P08453 (p94-p113) Immunogenic Deamidated DQ8 54 20 QTQQPQQPFPQQPQ QPFPQT 617 gamma-gliadin Predicted gamma-gliadin Immunogenic Native DQ8 (DQ2/8) 22 14 PFPQLQQPQQPFPQ 618 gamma-gliadin gamma-I gliadin 1206 Immunogenic Native DQ2  2 21 YQQLPQPQQPQQSF PQQQRPF 619 gamma-gliadin gamma-type gliadin of GDB2_WHEAT (SwissProt P08453) Immunogenic Deamidated DQ2 24 20 QQLPQPQQPQQSFP (p134-p153) QQQRPF 620 gamma-gliadin Glia-gamma1 epitope Immunogenic Native DQ2  9 17 QPQQPQQSFPQQQR PFI 621 gamma-gliadin Glia-gamma1 (p138-p153) Immunogenic Native DQ2 19 16 QPQQPQQSFPQQQR PF 622 gamma-gliadin Glia-gamma1 (p139-p153) Immunogenic Native DQ2  8 15 PQQPQQSFPQQQRP F 623 gamma-gliadin Glia-gamma1 (p139-p153; E148) Immunogenic Deamidated DQ2  8 15 PQQPQQSFPEQQRP F 624 gamma-gliadin Glia-gamma1 (p139-p153; E140 and E148) Immunogenic Deamidated DQ2  8 15 PEQPQQSFPEQQRP F 625 gamma-gliadin Glia-gamma1 (p139-p153; E148 and E150) Immunogenic Deamidated DQ2  8 15 PQQPQQSFPEQERP F 626 gamma-gliadin Glia-gamma1 (p139-p153; E140, E148 and E150) Immunogenic Deamidated DQ2  8 15 PEQPQQSFPEQERP F 627 gamma-gliadin gamma-I, gamma-Gliadin (p139-p152) Immunogenic Native DQ2 43, 25 14 PQQPQQSFPQQQRP 628 gamma-gliadin gamma-Gliadin (p139-p152; E140, E148 and E150) E Immunogenic Deamidated DQ2 43, 25 14 PEQPQQSFPEQERP residues in the gliadin peptides are introduced to mimic the deamidation mediated by tissue transglutaminase. 629 gamma-gliadin gamma-gliadin (p139-p152; E148) Immunogenic Deamidated DQ2 55 14 PQQPQQSFPEQQRP 630 gamma-gliadin gamma-gliadin (p139-p152; E140 and E148) Immunogenic Deamidated DQ2  14 PEQPQQSFPEQQRP (DQ2.2 and DQ2.5)25 631 gamma-gliadin P-3 gamma-gliadin (p139-p152; K139, E140, E148 and Immunogenic Synthesised DQ2 56 14 KEQPQQSFPEQERP E150) as Deamidated 632 gamma-gliadin P-2 gamma-gliadin (p139-p152; K140, E148 and E150) Immunogenic Synthesised DQ2 56 14 PKQPQQSFPEQERP as Deamidated 633 gamma-gliadin P-1 gamma-gliadin (p139-p152; K141, E140, E148 and Immunogenic Synthesised DQ2 56 14 PEKPQQSFPEQERP E150) as Deamidated 634 gamma-gliadin P1 y-gliadin(p139-p152; K142, E140, E148 and E150) Immunogenic Synthesised DQ2 56 14 PEQKQQSFPEQERP as Deamidated 635 gamma-gliadin P2 y-gliadin (p139-p152; K143, E140, E148 and E150) Immunogenic Synthesised DQ2 56 14 PEQPKQSFPEQERP as Deamidated 636 gamma-gliadin P4 y-gliadin (p139-p152; K144, E140, E148 and E150) Immunogenic Synthesised DQ2 56 14 PEQPQQKFPEQERP as Deamidated 637 gamma-gliadin P9 gamma-gliadin (p139-p152; E140, E148 and K150) Immunogenic Synthesised DQ2 56 14 PEQPQQSFPEQKRP as Deamidated 638 gamma-gliadin P10 gamma-gliadin (p139-p152; K151, E140, E148 and Immunogenic Synthesised DQ2 56 14 PEQPQQSFPEQEKP E150) as Deamidated 639 gamma-gliadin P11 y-gliadin (p139-p152; K152, E140, E148 and E150) Immunogenic Synthesised DQ2 56 14 PEQPQQSFPEQERK as Deamidated 640 gamma-gliadin gamma-I epitope in native form Immunogenic Native DQ2 47 12 QPQQSFPQQQRP 641 gamma-gliadin Deamidated form of gamma-I epitope Immunogenic Deamidated DQ2 47 12 QPQQSFPEQQRP 642 gamma-gliadin Glia-alpha20 Immunogenic Native DQ2  9 16 QQSFPQQQRPFIQP SL 643 gamma-gliadin gamma-gliadin AAK84772 (p130-p149) Immunogenic Native DQ8 54 20 PQPQQPQLPFPQQP QQPFPQ 644 gamma-gliadin Predicted gamma-gliadin Immunogenic Native DQ8 (DQ2/8) 22 14 PFPQPQQPQQPFPQ 645 gamma-gliadin gamma-gliadin AAK84776 (p102-p121) Immunogenic Native DQ8 54 20 QQPLPQPQQPQQPF PQSQQP 646 gamma-gliadin gamma-gliadin AAK84772 (p121-p140) Immunogenic Native DQ8 54 20 QPQQPQQPFPQQQQ PLIQPY 647 gamma-gliadin Wheat peptide W35 Immunogenic Native DQ2 62 20 PQQPFPQQPQQQFP QPQQPQ 648 gamma-gliadin Wheat peptide W35 Immunogenic Native DQ2 62 12 PFPQQPQQQFPQ 649 gamma-gliadin Wheat peptide W31 Immunogenic Native DQ2 62 20 QPFPQLQQPQQPLP QPQQPQ 650 gamma-gliadin Wheat peptide W31 Immunogenic Native DQ2 62 12 QPFPQLQQPQQP 651 LMW glutenin Wheat peptide W15 LMW Immunogenic Native DQ2 62 20 SHIPGLERPWQQQP LPPQQT 652 LMW glutenin Wheat peptide W15 LMW Immunogenic Native DQ2 62 15 QGLERPWQQQPLPP Q 653 LMW glutenin Wheat peptide W15 LMW Immunogenic Deamidated DQ2 62 15 EGLERPWQEQPLPP Q 654 LMW glutenin Wheat peptide W15 LMW Immunogenic Native DQ2 62 12 LERPWQQQPLPP 655 LMW glutenin Wheat peptide W11 Immunogenic Deamidated DQ2 62 16 GQQAFPQPEQTFPH QG 656 LMW glutenin Wheat peptide W11 Immunogenic Native DQ2 62 15 QQAFPQPQQTFPHQ P 657 LMW glutenin Wheat peptide W11 Immunogenic Deamidated DQ2 62 15 EQAFPQPEQTFPHQ P 658 LMW glutenin Wheat peptide W11 Immunogenic Native DQ2 62 20 QAFPQPQQTFPHQP QQQFPQ 659 LMW glutenin Wheat peptide W11 Immunogenic Native DQ2 62 12 QAFPQPQQTFPH 660 LMW glutenin Wheat peptide W11 Immunogenic Deamidated DQ2 62 12 QAFPQPEQTFPH 661 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60) Immunogenic Native DQ2 19 15 QQPPFSQQQQQPLP or LMW Q 662 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52) Immunogenic Deamidated DQ2 19 15 QQPPFSEQQQQPLP or LMW Q 663 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E53) Immunogenic Deamidated DQ2 19 15 QQPPFSQEQQQPLP or LMW Q 664 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E55) Immunogenic Deamidated DQ2 19 15 QQPPFSQQQEQPLP or LMW Q 665 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E56) Immunogenic Deamidated DQ2 19 15 QQPPFSQQQQEPLP or LMW Q 666 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52 and 53) Immunogenic Deamidated DQ2 19 15 QQPPFSEEQQQPLP or LMW Q 667 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52 and 55) Immunogenic Deamidated DQ2 19 15 QQPPFSEQQEQPLP or LMW Q 668 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSEQQQEPLP or LMW Q 669 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E53 and 55) Immunogenic Deamidated DQ2 19 15 QQPPFSQEQEQPLP or LMW Q 670 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E53 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSQEQQEPLP or LMW Q 671 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E55 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSQQQEEPLP or LMW Q 672 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52, 53 and 55) Immunogenic Deamidated DQ2 19 15 QQPPFSEEQEQPLP or LMW Q 673 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52, 53 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSEEQQEPLP or LMW Q 674 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E53, 55 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSQEQEEPLP or LMW Q 675 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52, 55 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSEQQEEPLP or LMW Q 676 gamma-gliadin gluteninGlutenin-Glt-17 (p46-p60; E52, 53, 55 and 56) Immunogenic Deamidated DQ2 19 15 QQPPFSEEQEEPLP or LMW Q 693 gamma-gliadin CAUTION 100% matches to 5 microbial proteins and to Immunogenic Native DQ2 27 9 FSQQQQQPL or LMW lutenin wheat proteins Glutenin-Glt-17 (p50-p58) 694 gamma-gliadin Glutenin-Glt-17 (p50-p58; E52) Immunogenic Deamidated DQ2 27 9 FSEQQQQPL or LMW lutenin 695 gamma-gliadin CAUTION 100% matches to two Pinus proteins/not wheat Immunogenic Deamidated DQ2 27 9 FSQEQQQPL or LMW lutenin Glutenin-Glt-17 (p50-p58; E53) 696 gamma-gliadin Glutenin-Glt-17 (p50-p58; E55) Immunogenic Deamidated DQ2 27 9 FSQQQEQPL or LMW lutenin 697 gamma-gliadin Glutenin-Glt-17 (p50-p58; E52 and E53) Immunogenic Deamidated DQ2 27 9 FSEEQQQPL or LMW lutenin 698 gamma-gliadin Glutenin-Glt-17 (p50-p58; E52 and E55) Immunogenic Deamidated DQ2 27 9 FSEQQEQPL or LMW lutenin 699 gamma-gliadin Glutenin-Glt-17 (p50-p58; E53 and E55) Immunogenic Deamidated DQ2 27 9 FSQEQEQPL or LMW lutenin 700 gamma-gliadin Glutenin-Glt-17 (p50-p58; E52, E53 and E55) Immunogenic Deamidated DQ2 27 9 FSEEQQEPL or LMW lutenin 701 gamma-gliadin Glutenin-17 epitope homolog Immunogenic Native DQ2 8, 19 15 QQPPFSQQQQPVLP or LMW lutenin Q 702 gamma-gliadin Glutenin-17 epitope homolog in Deamidated form Immunogenic Deamidated DQ2  8 15 QQPPFSEQQQPVLP or LMW lutenin Q 703 gamma-gliadin Glutenin-17 epitope homolog in Deamidated form Immunogenic Deamidated DQ2  8 15 QQPPFSQQEQPVLP or LMW lutenin Q 704 gamma-gliadin Glutenin-17 epitope homolog in Deamidated form Immunogenic Deamidated DQ2  8 15 QQPPFSEQEQPVLP or LMW lutenin Q 705 gamma-gliadin LMW T cell epitope Immunogenic Deamidated DQ2 62 15 EQPPFSEQEQPVLP or LMW lutenin Q 706 Glut-L1 CAUTION 100% match to a fungal protien Hebeloma sp. & Immunogenic Native DQ2.2 44, 90 9 PFSQQQQPV many wheat Glt-17 (Van) 707 glut-L1 Glt-17 (Van) Immunogenic Deamidated DQ2.2 44, 90 9 PFSEQQQPV 708 glut-L1 Glt-17 (Van) Immunogenic Deamidated DQ2.2 44, 90 9 PFSQQEQPV 709 glut-L1 Glt-17 (Van) Immunogenic Deamidated DQ2.2 44, 90 9 PFSEQEQPV 710 gamma-gliadin Wheat peptide W12 Immunogenic Native DQ2 62 20 CKVFLQQQCSPVAM or LMW lutenin PQRLAR 711 gamma-gliadin Wheat peptide W12 Immunogenic Native DQ2 62 16 LQQQCSPVAMPQRL or LMW lutenin AR 712 gamma-gliadin Wheat peptide W05 Immunogenic Native DQ2 62 20 PQQQQPFPQPQQPF or LMW-lutenin SQQPQQ 713 gamma-gliadin Wheat peptide W05 Immunogenic Deamidated DQ2 62 20 PQQQQPFPQPEQPF or LMW-lutenin SQQPQQ 714 gamma-gliadin Wheat peptide W05 Immunogenic Native DQ2 62 12 QPFPQPQQPFSQ or LMW-lutenin 715 gamma-gliadin Wheat peptide W05 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPFSQ or LMW-lutenin 716 gamma-gliadin Wheat peptide W17 Immunogenic Native DQ2 62 20 QQPFPQPQQPQLPF or LMW-lutenin PQQPQQ 717 gamma-gliadin Wheat peptide W17 Immunogenic Native DQ2 62 15 QPFPQPQQPQLPFP or LMW-lutenin Q 718 gamma-gliadin Wheat peptide W17 Immunogenic Deamidated DQ2 62 15 EPFPQPEQPELPFP or LMW-lutenin Q 719 gamma-gliadin Wheat peptide W17 Immunogenic Native DQ2 62 12 QPFPQPQQPQLP or LMW-lutenin 720 LMW glutenin GLT/GLIA homologue peptide 12 Immunogenic Native DQ2 19 15 QQPPFSQQQQPPFS Q 721 LMW glutenin LMW glutenin-glt-156 (p40-p59) Immunogenic Native DQ2 19 20 QQQQPPFSQQQQSP FSQQQQ 722 LMW glutenin LMW glutenin-glt-156 (p40-p59; E48) Immunogenic Deamidated DQ2 19 20 QQQQPPFSEQQQSP FSQQQQ 723 LMW glutenin LMW glutenin-glt-156 (p40-p59; E51) Immunogenic Deamidated DQ2 19 20 QQQQPPFSQQQESP FSQQQQ 724 LMW glutenin LMW glutenin-glt-156 (p40-p59; E48 and E51) Immunogenic Deamidated DQ2 19 20 QQQQPPFSEQQESP FSQQQQ 725 LMW glutenin Homolog of Deamidated Glt-156 minimal epitope Immunogenic Deamidated DQ2 20 15 QQQQPPFSEEQESP (p40-p59) Y 726 LMW glutenin Homolog of Deamidated Glt-156 minimal epitope Immunogenic Deamidated DQ2 20 15 QQQQPPFSEEQESP (p40-p59) L 727 LMW glutenin Deamidated Glt-156 minimal epitope (p40-p59) Immunogenic Deamidated DQ2 20 15 QQQQPPFSEEQESP F 728 LMW glutenin Glt-156 minimal epitope (p41-p55) Immunogenic Deamidated DQ2 20 15 QQQPPFSEEQESPF S 729 LMW glutenin Glt-156 minimal epitope in considered native form Immunogenic Native DQ2 20 15 QQPPFSQQQQSPFS Q 730 LMW glutenin Glt-156 minimal epitope in considered Deamidated Immunogenic Deamidated DQ2 20 15 QQPPFSEEQESPFS form Q 731 LMW glutenin GLT/GLIA homologue peptide 4 Immunogenic Native DQ2 19 12 QQPPFSQQQQSP 732 LMW glutenin Glt-156 minimal epitope Immunogenic Deamidated DQ2 20 15 QPPFSEEQESPFSQ Q 733 LMW glutenin LMW glutenin-glt-156 (p40-p59) Immunogenic Native DQ2 16 14 QPPFSQQQQSPFSQ 734 LMW glutenin Glt-156 minimal epitope in considered native form Immunogenic Native DQ2 20 15 PPFSQQQQSPFSQQ Q 735 LMW glutenin Glt-156 minimal epitope in considered Deamidated form Immunogenic Deamidated DQ2 20 15 PPFSEEQESPFSQQ Q 736 LMW glutenin Glt-156 minimal epitope in considered native form Immunogenic Native DQ2 20 15 PFSQQQQSPFSQQQ Q 737 LMW glutenin Glt-156 minimal epitope in considered Deamidated form Immunogenic Deamidated DQ2 20 15 PFSEEQESPFSQQQ Q 738 LMW glutenin LMW glutenin-glt-156 (p45-p54) minimal epitope Immunogenic Native DQ2 19, 20 10 PFSQQQQSPF 739 LMW glutenin LMW glutenin-glt-156 (p45-p54; E48) minimal epitope Immunogenic Deamidated DQ2 19, 20 10 PFSEQQQSPF 740 LMW glutenin LMW glutenin-glt-156 (p45-p54; E49) minimal epitope Immunogenic Deamidated DQ2 20 10 PFSQEQQSPF 741 LMW glutenin LMW glutenin-glt-156 (p45-p54; E51) minimal epitope Immunogenic Deamidated DQ2 19, 20 10 PFSQQQESPF 742 LMW glutenin LMW glutenin-glt-156 (p45-p54; E48 and E51) minimal Immunogenic Deamidated DQ2 19, 20 10 PFSEQQESPF epitope 743 LMW glutenin LMW glutenin-glt-156 (p45-p54; E48 and E49) minimal Immunogenic Deamidated DQ2 20 10 PFSEEQQSPF epitope 745 LMW glutenin LMW glutenin-glt-156 (p45-p54; E49 and E49) minimal Immunogenic Deamidated DQ2 20 10 PFSQEQESPF epitope 746 LMW glutenin LMW glutenin-glt-156 (p45-p54; E48, E49 and E51) Immunogenic Deamidated DQ2 20 10 PFSEEQESPF minimal epitope 747 glut-L2 LMW glutenin-glt-156 (p46-p54) Immunogenic Native DQ2.5 19, 90, 27 9 FSQQQQSPF 748 glut-L2 LMW glutenin-glt-156 (p46-p54; E48) Immunogenic Deamidated DQ2.5 19, 90, 27 9 FSEQQQSPF 749 glut-L2 LMW glutenin-glt-156 (p46-p54; E51) Immunogenic Deamidated DQ2.5 19, 90, 27 9 FSQQQESPF 750 glut-L2 LMW glutenin-glt-156 (p46-p54; E48 and E51) Immunogenic Deamidated DQ2.5 19, 90, 27 9 FSEQQESPF 751 LMW glutenin GLT/GLIA homologue peptide 13 Immunogenic Native DQ2 19 15 QQPPFSQQQQPQFS Q 752 LMW glutenin Gluten peptide E19025 Immunogenic Native DQ2 (DQ2.5) 61 19 SHQQQPFPQQPYPQ QPYPS 753 gamma-gliadin 14-mer-2 gamma-Glia (p173-p186) Immunogenic Native DQ2 57 14 PQQPFPSQQQQPLI 754 gamma-gliadin 14-mer-2 gamma-Glia (p173-p186) in Deamidated form Immunogenic Deamidated DQ2 57 14 PQQPFPSQQEQPLI 755 LMW glutenin GLT/GLIA homologue peptide 17 Immunogenic Native DQ2 19 15 QQPPFSQQQQPILP Q 756 LMW glutenin Glt-156 homolog Immunogenic Native DQ2 21 14 QPPFSQQQQPILPQ 757 LMW glutenin Glt-156 homolog in Deamidated form Immunogenic Deamidated DQ2 21 14 QPPFSEQEQPILPQ 762 LMW glutenin GLT/GLIA homologue peptide 16 Immunogenic Native DQ2 19 15 QQPPFSQQQQQPIL L 763 LMW glutenin Glt-156 homolog Immunogenic Native DQ2 21 14 QPPFSQQQQQPILL 764 LMW glutenin Glt-156 homolog in Deamidated form Immunogenic Deamidated DQ2 21 14 QPPFSEEQEQPILL 765 HMW glutenin Wheat peptide W21 HMW Immunogenic Native DQ2 62 20 QGQQGYYPISPQQS GQGQQP 766 HMW glutenin Wheat peptide W21 HMW Immunogenic Native DQ2 62 16 QGQQGYYPISPQQS GQ 767 HMW glutenin Wheat peptide W21 HMW Immunogenic Deamidated DQ2 62 16 EGQQGYYPISPQQS GQ 768 HMW glutenin Naturally occurring glutenins p722-p736 (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQAGYYPTSPQQSG of glt04) Q 769 HMW glutenin Naturally occurring glutenins p722-p736 (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSPQQPG of glt04) Q 770 HMW glutenin Naturally occurring glutenins p722-p736 (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPISPQQSG of glt04) Q 771 HMW glutenin Wheat peptide W29 Immunogenic Native DQ2 62 20 GQGQSGYYPTSPQQ SGQEAT 772 HMW glutenin Wheat peptide W29 Immunogenic Native DQ2 62 16 GQGQSGYYPTSPQQ SG 773 HMW glutenin Wheat peptide W24 HMW Immunogenic Native DQ2 62 20 PGQGQSGYYPTSPQ QSGQKQ 774 HMW glutenin Wheat peptide W24 HMW Immunogenic Native DQ2 62 16 PGQGQSGYYPTSPQ QS 775 HMW glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQSGYYPTSPQQSG of glt04) Q 776 HMW glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPISPQQLG of glt04) Q 777 HMW glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQLGYYPTSPQQSG of glt04) Q 778 HMW glutenin Wheat peptide W22 HMW Immunogenic Native DQ2 62 20 LQPGQGQPGYYPTS PQQIGQ 779 HMW glutenin Wheat peptide W22 HMW Immunogenic Native DQ2 62 16 QGQPGYYPTSPQQI GQ 780 HMW glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQPGYYPTSPQQIG of glt04) Q 781 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQPGYYPTSPQQPG of glt04) Q 782 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQPGYYPTSPQQSG of glt04) Q 783 HMW-Glutenin Naturally occurring glutenins p722-p736 (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSLQQPG of glt04) Q 784 HMW-Glutenin HMW glutenin-glt04 (p707-p742) Immunogenic Native DQ8 (DQ2/8)  7 36 SGQGQRPGQWLQPG   QGQQGYYPTSPQQS GQGQQLGQ 785 HMW-Glutenin HMW glutenin-glt04 (p719-p736) Immunogenic Native DQ8 (DQ2/8)  7 18 PGQGQQGYYPTSPQ QSGQ 786 HMW-Glutenin glt04 (p722-p736) Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSPQQSG Q 787 HMW-Glutenin glt04 (p722-p735) Immunogenic Native DQ8 (DQ2/8)  7 14 GQQGYYPTSPQQSG 788 HMW-Glutenin glt04 (p722-p734) Immunogenic Native DQ8 (DQ2/8)  7 13 GQQGYYPTSPQQS 789 HMW-Glutenin glt04 (p723-p736) Immunogenic Native DQ8 (DQ2/8)  7 14 QQGYYPTSPQQSGQ 790 HMW-Glutenin glt04 (p723-p735) Immunogenic Native DQ8 (DQ2/8)  7 13 QQGYYPTSPQQSG 791 HMW-Glutenin glt04 (p723-p735; E724) Immunogenic Deamidated DQ8 (DQ2/8) 7, 83 13 QEGYYPTSPQQSG 792 HMW-Glutenin glt04 (p723-p734) Immunogenic Native DQ8 (DQ2/8) 52 12 QQGYYPTSPQQS 793 HMW-Glutenin glt04 (p724-p735) Immunogenic Native DQ8 (DQ2/8)  7 12 QGYYPTSPQQSG 794 glut-H1 HMW glutenin (p724-p734) Immunogenic Native DQ8, DQ8.5 7, 76, 27 11 QGYYPTSPQQS 797 HMW-Glutenin glt04 (p725-p735) Immunogenic Native DQ8 (DQ8.5) 7, 83 11 GYYPTSPQQSG 798 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPISPQQPG of glt04) Q 799 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSPQQSP of glt04) Q 800 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSPQQLG of glt04) Q 801 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSPQHPG of glt04) Q 802 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQPGYYPTSPLQSG of glt04) Q 803 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQHGYYPTSPQLSG of glt04) Q 804 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSPQQPP of glt04) Q 805 HMW-Glutenin Naturally occurring glutenins (p722-p736) (homolog Immunogenic Native DQ8 (DQ2/8)  7 15 GQQGYYPTSVQQPG of glt04) Q 806 Hordein Barley peptide B04, B17 Immunogenic Native DQ2 62 20 QPQQPQPFPQQPVP QQPQPY 807 Hordein Barley peptide B17 Immunogenic Native DQ2 62 16 PQQPQPFPQQPVPQ QP 808 Hordein Barley peptide B04 Immunogenic Native DQ2 62 12 PQQPVPQQPQPY 809 Hordein Barley peptide B05, B08 in native form Immunogenic Native DQ2 62 20 PQPFPQQPIPQQPQ PYPQQP 810 Hordein Barley peptide B05, B08 in Deamidated form Immunogenic Deamidated DQ2 62 20 PQPFPQQPIPEQPQ PYPQQP 811 Hordein Barley peptide B05 Immunogenic Native DQ2 62 16 PQPFPQQPIPQQPQ PY 812 Hordein Barley peptide B05 Immunogenic Deamidated DQ2 62 16 PQPFPQQPIPEQPQ PY 813 Hordein Barley peptide B06 Immunogenic Native DQ2 62 20 QQPQPFSQQPIPQQ PQPYPQ 814 Hordein Barley peptide B06 Immunogenic Deamidated DQ2 62 20 QQPQPFSQQPIPEQ PQPYPQ 815 Hordein Barley peptide B06 Immunogenic Native DQ2 62 12 SQQPIPQQPQPY 816 Hordein Barley peptide B06 Immunogenic Deamidated DQ2 62 12 SQQPIPEQPQPY 817 Hordein Barley peptide B18 Immunogenic Native DQ2 62 20 QPQPFPQQPIPLQP HQPYTQ 818 Hordein Barley peptide B18 Immunogenic Native DQ2 62 12 QPQPFPQQPIPL 819 Hordein Barley peptide B13 Immunogenic Native DQ2 62 20 PQPYPQQPQPFPQQ PPFCQQ 820 Hordein Barley peptide B13 Immunogenic Native DQ2 62 16 PQPYPQQPQPFPQQ PP 821 Hordein Barley peptide B09, B12, B30 Immunogenic Native DQ2 62 20 QQPFPQQPFPQQPQ PYPQQP 822 Hordein Barley peptide B09 Immunogenic Native DQ2 62 16 QQPFPQQPFPQQPQ PY 823 Hordein Barley peptide B30 Immunogenic Native DQ2 62 11 QQPFPQQPFPQ 824 Hordein Barley peptide B12 Immunogenic Native DQ2 62 16 PQQPFPQQPQPYPQ QP 825 Hordein Barley peptide B11 Immunogenic Native DQ2 62 20 QPQPYPQQPQPYPQ QPFQPQ 826 Hordein Barley peptide B11 Immunogenic Native DQ2 62 12 QPQPYPQQPQPY 827 gamma-gliadin Glu-21 in considered native form Immunogenic Native DQ2 19 21 QPQPFPQQSEQSQQ or LMW PFQPQPF glutenin 828 Hordein Barley peptide B03 Immunogenic Native DQ2 62 20 QPQQPFPQPQQPIP YQPQQP 829 Hordein Barley peptide B03 Immunogenic Deamidated DQ2 62 16 GQQPFPQPEQPIPY QG 830 Hordein Barley peptide B03 Immunogenic Native DQ2 62 12 QPFPQPQQPIPY 831 Hordein Barley peptide B03 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPIPY 832 Hordein Barley peptide B02 Immunogenic Native DQ2 62 20 WQPQQPFPQPQQPF PLQPQQ 833 Hordein Barley peptide B02 Immunogenic Deamidated DQ2 62 20 WQPQQPFPQPEQPF PLQPQQ 834 Hordein Barley peptide B02 Immunogenic Deamidated DQ2 62 16 GQQPFPQPEQPFPL QG 835 Hordein Barley peptide B02 Immunogenic Native DQ2 62 12 QPFPQPQQPFPL 836 Hordein Barley peptide B02 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPFPL 837 Hordein Barley peptide B19 Immunogenic Native DQ2 62 20 LPRPQQPFPWQPQQ PFPQPQ 838 Hordein Barley peptide B26 Immunogenic Native DQ2 62 20 QPQQPFPLQPQQPF PWQPQQ 839 Hordein Barley peptide B26 Immunogenic Native DQ2 62 12 PFPLQPQQPFPW 840 Hordein Barley peptide B29 Immunogenic Native DQ2 62 20 QPQQPFSFSQQPQQ PFPLQP 841 Hordein Barley peptide B29 Immunogenic Deamidated DQ2 62 16 GFSFSQQPEQPFPL QG 842 Hordein Barley peptide B14 Immunogenic Native DQ2 62 20 FQQPQQSYPVQPQQ PFPQPQ 843 Hordein Barley peptide B14 Immunogenic Deamidated DQ2 62 16 GQSYPVQPEQPFPQ PG 844 Hordein Barley peptide B14 Immunogenic Native DQ2 62 12 SYPVQPQQPFPQ 845 Hordein Barley peptide B14 Immunogenic Deamidated DQ2 62 12 SYPVQPEQPFPQ 846 Hordein Barley peptide B29 Immunogenic Native DQ2 62 12 SFSQQPQQPFPL 847 Hordein Barley peptide B29 Immunogenic Deamidated DQ2 62 12 SFSQQPEQPFPL 848 omega-gliadin Wheat peptide W19 Immunogenic Deamidated DQ2 62 16 GQPFPWQPEQPFPQ PG 849 Hordein Barley peptide B15 Immunogenic Native DQ2 62 20 YPQQPQPFPQQPIP QQPQPY 850 Hordein Barley peptide B15 Immunogenic Native DQ2 62 12 QPQPFPQQPIPQ 851 hor-3 Hor-I Immunogenic Native DQ2.5 88, 62 9 PIPQQPQPY 852 hor-3 Hor-I Immunogenic Deamidated DQ2.5 88, 62 9 PIPEQPQPY 853 Hordein Barley peptide B16 Immunogenic Native DQ2 62 20 QQQPFPQQPIPQQP QPYPQQ 854 Hordein Barley peptide B16 Immunogenic Native DQ2 62 11 QQPFPQQPIPQ 855 Hordein Barley peptide B08 Immunogenic Deamidated DQ2 62 16 PQQPIPQQPQPYPQ QP 856 Hordein Barley peptide B08 Immunogenic Deamidated DQ2 62 16 PQQPIPEQPQPYPQ QP 857 Hordein Barley peptide B08 Immunogenic Native DQ2 62, 86 16 QPQQPIPQQPQPYP QQ 858 Hordein Barley peptide B08 Immunogenic Deamidated DQ2 62, 86 16 EPEQPIPEQPQPYP QQ 859 Hordein Hordein core epitope in native form Immunogenic Native DQ2 27 9 FPPQQPFPQ 860 Hordein Hordein core epitope in demainated form Immunogenic Deamidated DQ2 27 9 FPPEQPFPQ 861 Hordein Barley peptide B21, 625 Immunogenic Native DQ2 62 20 PFPQQPQQPFPQPQ QPFRQQ 862 Hordein Barley peptide B21 Immunogenic Deamidated DQ2 62 20 PFPQQPQQPFPQPE QPFRQQ 863 Hordein alpha9-Hordein in native form Immunogenic Native DQ2  8 14 PQQPFPQPQQPFRQ 864 Hordein alpha9-Hordein in Deamidated form Immunogenic Deamidated DQ2  8 14 PQQPFPQPEQPFRQ 865 Hordein Barley peptide B21 Immunogenic Native DQ2 62 13 QQPFPQPQQPFRQ 866 Hordein Barley peptide B21 Immunogenic Deamidated DQ2 62 13 QQPFPQPEQPFRQ 867 hor-1 Hordein/Secalin Immunogenic Native DQ2.5 88, 90, 8, 9 PFPQPQQPF 27, 62 868 hor-1 Hordein/Secalin Immunogenic Deamidated DQ2.5 88, 90, 8, 9 PFPQPEQPF 27, 62 869 Hordein Barley peptide B22 Immunogenic Native DQ2 62 20 PQQPFQPQQPFPQQ TIPQQP 870 Hordein Barley peptide B22 Immunogenic Native DQ2 62 12 QQPFQPQQPFPQ 871 Hordein Barley peptide B27 Immunogenic Native DQ2 62 20 TFPPSQQPNPLQPQ QPFPLQ 872 Hordein Barley peptide B27 Immunogenic Native DQ2 62 13 PNPLQPQQPFPLQ 873 Hordein Barley peptide B23, 624 Immunogenic Native DQ2 62 20 NPLQPQQPFPLQPQ PPQQPF 874 Hordein Barley peptide B23 Immunogenic Native DQ2 62 16 NPLQPQQPFPLQPQ PP 875 Hordein Barley peptide B24 Immunogenic Native DQ2 62 16 PLQPQQPFPLQPQP PQ 876 Hordein Barley peptide B10 Immunogenic Native DQ2 62 20 PQQPQQPFPQPQQP FSWQPQ 877 Hordein Barley peptide B10 Immunogenic Deamidated DQ2 62 20 PQQPQQPFPQPEQP FSWQPQ 878 Hordein Barley peptide B10 Immunogenic Native DQ2 62 12 QPFPQPQQPFSW 879 Hordein Barley peptide B10 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPFSW 880 Hordein Barley peptide B28 Immunogenic Native DQ2 62 20 PQQTIPQQPQQPFP LQPQQP 881 Hordein Barley peptide B28 Immunogenic Native DQ2 62 12 TIPQQPQQPFPL 882 Hordein Barley peptide B20 Immunogenic Native DQ2 62 20 QQPFPLQPQQPFPQ PQPFPQ 883 gamma-gliadin Wheat peptide W26 Immunogenic Deamidated DQ2 62 16 GQPFPLQPEQPFPQ PG 884 gamma-gliadin Wheat peptide W26 Immunogenic Deamidated DQ2 62 12 PFPLQPEQPFPQ 885 gamma-hordein Barley peptide B07 Immunogenic Native DQ2 62 20 QSQQQFPQPQQPFP QQPQQP 886 gamma-hordein alpha2-Hordein in native form Immunogenic Native DQ2  8 14 QQFPQPQQPFPQQP 887 gamma-hordein alpha2-Hordein in Deamidated form Immunogenic Deamidated DQ2  8 14 QEFPQPQQPFPQQP 888 gamma-hordein alpha2-Hordein in Deamidated form Immunogenic Deamidated DQ2  8 14 QQFPQPEQPFPQQP 889 gamma-hordein alpha2-Hordein in Deamidated form Immunogenic Deamidated DQ2  8 14 QEFPQPEQPFPQQP 890 gamma-hordein Barley peptide B07 Immunogenic Native DQ2 62 12 QQFPQPQQPFPQ 891 hor-2 Hordein/Secalin Immunogenic Native DQ2.5 90, 8, 27 9 PQPQQPFPQ 892 hor-2 Hordein/Secalin Immunogenic Deamidated DQ2.5 90, 8, 27 9 PQPEQPFPQ 893 Secalin Rye peptide R05 Immunogenic Deamidated DQ2 62 16 GQPAPIQPEQPFPQ QG 894 Secalin Rye peptide R05, R26 Immunogenic Native DQ2 62 20 PAPIQPQQPFPQQP QQPFPQ 895 Secalin Rye peptide R05 Immunogenic Deamidated DQ2 62 12 PAPIQPEQPFPQ 896 Secalin Rye peptide R05 Immunogenic Native DQ2 62 12 PAPIQPQQPFPQ 897 Secalin Rye peptide R12 Immunogenic Native DQ2 62 20 FPQQPQQPFPQPQQ QLPLQP 898 Secalin Rye peptide R12 Immunogenic Deamidated DQ2 62 16 GQQPFPQPEQELPL QG 899 Secalin Rye peptide R12 Immunogenic Native DQ2 62 12 QPFPQPQQQLPL 900 Secalin Rye peptide R12 Immunogenic Deamidated DQ2 62 12 QPFPQPEQELPL 901 Secalin Rye peptide R29 Immunogenic Native DQ2 62 20 PTPIQPQQPFPQRP QQPFPQ 902 Secalin Rye peptide R29 Immunogenic Native DQ2 62 12 PFPQRPQQPFPQ 903 Secalin gamma1-Secalin in native form Immunogenic Native DQ2 90, 27 9 PQQSFPQQP 904 Secalin gamma1-Secalin in Deamidated form Immunogenic Deamidated DQ2 90, 27 9 PQQSFPEQP 905 Secalin Rye peptide R10 Immunogenic Native DQ2 62 20 FPLQPQQPFPQQPE QIISQQ 906 Secalin Rye peptide R10 Immunogenic Native DQ2 62 12 PFPQQPEQIISQ 907 Secalin Rye peptide R25 Immunogenic Native DQ2 62 20 FPQQPEQIISQQPQ QPFPLQ 908 Secalin Rye peptide R25 Immunogenic Native DQ2 62 15 PEQIISQQPQQPFP L 909 Secalin Rye peptide R22 Immunogenic Native DQ2 62 20 PQQLFPLPQQPFPQ PQQPFP 910 Secalin Rye peptide R22 Immunogenic Native DQ2 62 11 LFPLPQQPFPQ 911 Secalin alpha9-Secalin in native form Immunogenic Native DQ2  8 14 PQQPFPQPQQPFPQ 912 Secalin alpha9-Secalin in Deamidated form Immunogenic Deamidated DQ2  8 14 PEQPFPQPQQPFPQ 913 Secalin alpha9-Secalin in Deamidated form Immunogenic Deamidated DQ2  8 14 PQQPFPQPEQPFPQ 914 Secalin alpha9-Secalin in Deamidated form Immunogenic Deamidated DQ2  8 14 PEQPFPQPEQPFPQ 915 Secalin alpha2-Secalin in native form Immunogenic Native DQ2  8 14 QPFPQPQQPFPQSQ 916 Secalin alpha2-Secalin in Deamidated form Immunogenic Deamidated DQ2  8 14 QPFPQPEQPFPQSQ 917 gamma-secalin Rye peptide R21 Immunogenic Native DQ2 62 20 NMQVGPSGQVEWPQ QQPLPQ 918 gamma-secalin Rye peptide R21 Immunogenic Native DQ2 62 16 GMQVGPSGEVEWPQ QG 919 gamma-secalin Rye peptide R21 Immunogenic Native DQ2 62 12 QVGPSGQVEWPQ 920 gamma-secalin Rye peptide R21 Immunogenic Native DQ2 62 12 QVGPSGEVEWPQ 921 gamma-secalin Rye peptide R13, R28 Immunogenic Native DQ2 62 20 SPQPQQPYPQQPFP QQPQQP 922 gamma-secalin Rye peptide R13 Immunogenic Deamidated DQ2 62 16 GQPEQPYPEQPFPQ QG 923 gamma-secalin Rye peptide R13 Immunogenic Native DQ2 62 12 PQQPYPQQPFPQ 924 gamma-secalin Rye peptide R13 Immunogenic Deamidated DQ2 62 12 PEQPYPEQPFPQ 925 gamma-secalin Rye peptide R23 Immunogenic Native DQ2 62 20 PQTQQPQQPFPQPQ QPQQLF 926 gamma-secalin Rye peptide R23 Immunogenic Native DQ2 62 12 PQTQQPQQPFPQ 927 gamma-secalin Rye peptide R27 Immunogenic Native DQ2 62 20 PQEPQQLFPQSQQP QQPFPQ 928 gamma-secalin Rye peptide R27 Immunogenic Native DQ2 62 12 PQSQQPQQPFPQ 929 gamma-secalin Rye peptide R17 Immunogenic Native DQ2 62 20 QTQQSIPQPQQPFP QPQQPF 930 gamma-secalin Rye peptide R17 Immunogenic Native DQ2 62 12 QSIPQPQQPFPQ 931 gamma-secalin Rye peptide R02 Immunogenic Native DQ2 62 20 SIPQPQQPFPQPQQ PFPQSQ 932 gamma-secalin Rye peptide R02 Immunogenic Deamidated DQ2 62 20 SIPQPQQPFPQPEQ PFPQSQ 933 gamma-secalin Rye peptide R02 Immunogenic Deamidated DQ2 62 16 GQQPFPQPEQPFPQ SG 934 gamma-secalin Rye peptide R02 Immunogenic Deamidated DQ2 62 13 QPFPQPEQPFPQS 935 gamma-secalin Rye peptide R02 Immunogenic Native DQ2 62 12 QPFPQPQQPFPQ 936 gamma-secalin Rye peptide R02 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPFPQ 937 omega-Secalin Rye peptide R07 Immunogenic Native DQ2 62 20 QYSPYQPQQPFPQP QQPTPI 938 omega-Secalin Rye peptide R07 Immunogenic Deamidated DQ2 62 16 GQYSPYQPEQPFPQ PG 939 omega-Secalin Rye peptide R07 Immunogenic Native DQ2 62 12 YSPYQPQQPFPQ 940 omega-Secalin Rye peptide R07 Immunogenic Deamidated DQ2 62 12 YSPYQPEQPFPQ 941 omega-Secalin Rye peptide R03 Immunogenic Deamidated DQ2 62 16 GQQPFPQPEQPTPI QG 942 omega-Secalin Rye peptide R03, R04 Immunogenic Native DQ2 62 20 QPFPQPQQPTPIQP QQPFPQ 943 omega-Secalin Rye peptide R03 Immunogenic Deamidated DQ2 62 12 QPFPQPEQPTPI 944 omega-Secalin Rye peptide R03 Immunogenic Native DQ2 62 12 QPFPQPQQPTPI 945 omega-Secalin Rye peptide R04 Immunogenic Deamidated DQ2 62 16 GQPTPIQPEQPFPQ QG 946 omega-Secalin Rye peptide R04 Immunogenic Native DQ2 62 12 PTPIQPQQPFPQ 947 omega-Secalin Rye peptide R04 Immunogenic Deamidated DQ2 62 12 PTPIQPEQPFPQ 948 omega-Secalin Rye peptide R01, R09 Immunogenic Native DQ2 62 20 QQLPLQPQQPFPQP QQPIPQ 949 omega-Secalin Rye peptide R09 Immunogenic Native DQ2 62 12 QLPLQPQQPFPQ 950 omega-Secalin Rye peptide R01 Immunogenic Native DQ2 62 12 QPFPQPQQPIPQ 951 omega-Secalin Sec-gamma1 Immunogenic Native DQ2  8 14 PQQPQQSFPQQPQR 952 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PEQPQQSFPQQPQR 953 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PQQPEQSFPQQPQR 954 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PQQPQQSFPEQPQR 955 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PEQPEQSFPQQPQR 956 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PEQPQQSFPEQPQR 957 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PQQPEQSFPEQPQR 958 omega-Secalin Sec-gamma1 in Deamidated form Immunogenic Deamidated DQ2  8 14 PEQPEQSFPEQPQR 959 omega-Secalin Rye peptide R20 Immunogenic Native DQ2 62 20 EQIISQQPFPLQPQ QPFSQP 960 omega-Secalin Rye peptide R20 Immunogenic Native DQ2 62 12 PFPLQPQQPFSQ 961 omega-Secalin Rye peptide R6 Immunogenic Deamidated DQ2 62 16 GQPQQPFPEQPEQI IG 962 omega-Secalin Rye peptide R06, R11, R16 Immunogenic Native DQ2 62 20 PQQPFPQQPEQIIP QQPQQP 963 omega-Secalin Rye peptide R6 Immunogenic Native DQ2 62 12 PQQPFPQQPEQI 964 omega-Secalin Rye peptide R6 Immunogenic Deamidated DQ2 62 12 PQQPFPEQPEQI 965 omega-Secalin Rye peptide R11 Immunogenic Native DQ2 62 16 QQPFPQQPEQIIPQ QP 966 omega-Secalin Rye peptide R11 Immunogenic Deamidated DQ2 62 16 EQPFPEQPEQIIPQ QP 967 omega-Secalin Rye peptide R11 Immunogenic Deamidated DQ2 62 16 GQPFPQQPEQIIPQ QG 968 omega-Secalin Rye peptide R11 Immunogenic Deamidated DQ2 62 12 PFPQQPEQIIPQ 969 omega-Secalin Rye peptide R16 Immunogenic Native DQ2 62 12 PEQIIPQQPQQP 970 omega-Secalin Rye peptide R08 Immunogenic Native DQ2 62 20 SQQPQRPQQPFPQQ PQQIIP 971 omega-Secalin Rye peptide R08 Immunogenic Native DQ2 62 13 RPQQPFPQQPQQI 972 omega-Secalin Rye peptide R15 Immunogenic Native DQ2 62 20 QPQQI1PQQPQQPF PLQPQQ 973 omega-Secalin Rye peptide R15 Immunogenic Native DQ2 62 12 IIPQQPQQPFPL 974 omega-Secalin Rye peptide R14, R19 Immunogenic Native DQ2 62 20 QQPQQPFPLQPQQP VPQQPQ 975 omega-Secalin Rye peptide R14 Immunogenic Native DQ2 62 16 QQPQQPFPLQPQQP VP 976 omega-Secalin Rye peptide R19 Immunogenic Native DQ2 62 16 QPFPLQPQQPVPQQ PQ 977 omega-Secalin Rye peptide R18 Immunogenic Native DQ2 62 20 QQPFLLQPQQPFSQ PQQPFL 978 omega-Secalin Rye peptide R18 Immunogenic Native DQ2 62 11 FLLQPQQPFSQ 979 omega-Secalin Rye peptide R24 Immunogenic Native DQ2 62 20 SPQQPQLPFPQPQQ PFVVVV 980 omega-Secalin Rye peptide R24 Immunogenic Deamidated DQ2 62 20 SPQQPQLPFPQPEQ PFVVVV 981 omega-Secalin Rye peptide R24 Immunogenic Native DQ2 62 12 LPFPQPQQPFVV 982 omega-Secalin Rye peptide R24 Immunogenic Deamidated DQ2 62 12 LPFPQPEQPFVV 983 gamma-avenin Av-alpha9B in native form Immunogenic Native DQ2  8 14 QYQPYPEQQQPFVQ 984 gamma-avenin Av-alpha9B in Deamidated form Immunogenic Deamidated DQ2  8 14 QYQPYPEQEQPFVQ 985 gamma-avenin Homolog of oat avenin-derived T cell-stimulatory Immunogenic Deamidated DQ2 62 15 EYQPYPEQEQPILQ peptide in Deamidated form Q 986 gamma-avenin Homolog of oat avenin-derived T cell-stimulatory Immunogenic Native DQ2 62 15 QYQPYPQQQQPILQ peptide in native form Q 987 ave-1b gliadin alpha avenin-9 Immunogenic Native DQ2.5 90, 18, 8 9 PYPEQQQPF 988 ave-1b gliadin alpha avenin-9 Immunogenic Deamidated DQ2.5 90, 18, 8 9 PYPEQEQPF 989 Avenin T cell recognized Avenin epitope HPLC fraction 9 Immunogenic Native DQ2 18 31 TTTVQYDPSEQYQP YPEQQEPFVQQQPP FVQ 990 Avenin T cell recognized Avenin epitope HPLC fraction 4 Immunogenic Native DQ2 18 22 TTTVQYDPSEQYQP YPEQQEPF 991 Avenin T cell recognized Avenin epitope HPLC fraction 9 Immunogenic Native DQ2 18 28 TTTVQYNPSEQYQP YPEQQEPFVQQQPF 992 Avenin T cell recognized Avenin epitope HPLC fraction 9 Immunogenic Native DQ2 18 27 TTVQYNPSEQYQPY PEQQEPFVQQQPF 993 Avenin T cell recognized Avenin epitope HPLC fraction 9 Immunogenic Native DQ2 18 25 VQYNPSEQYQPYPE QQEPFVQQQPF 994 Avenin T cell recognized Avenin epitope HPLC fraction 3 Immunogenic Native DQ2 18 22 TTTVQYNPSEQYQP YPEQQEPF 995 Avenin T cell recognized Avenin epitope HPLC fraction 8 Immunogenic Native DQ2 18 29 TTTVQYDPSEQYQP YPEQQEPFVQQQQP F 996 Avenin T cell recognized Avenin epitope HPLC fraction 8 Immunogenic Native DQ2 18 30 TTTVQYNPSEQYQP YPEQQEPFVQQQQP FV 997 Avenin T cell recognized Avenin epitope HPLC fraction 9 Immunogenic Native DQ2 18 29 PSEQYQPYPEQQEP FVQQQQPFVQQQQP F 998 Avenin Avenin 1490 in native form Immunogenic Native DQ2 18 19 SEQYQPYPEQQEPF VQQQQ 999 Avenin Avenin 1490 in Deamidated form Immunogenic Deamidated DQ2 18 19 SEQYQPYPEQEEPF VQQQQ 1000 Avenin Av-alpha9A in native form Immunogenic Native DQ2  8 14 QYQPYPEQQEPFVQ 1001 Avenin Av-alpha9A in Deamidated form Immunogenic Deamidated DQ2  8 14 QYQPYPEQEEPFVQ 1002 Avenin Avenin 1505 Immunogenic Native DQ2 18 12 YQPYPEQQEPFV 1003 Avenin Avenin 1504 (deamidated form of Avenin 1505) Immunogenic Deamidated DQ2 18 12 YQPYPEQEEPFV 1004 ave-1 gliadin alpha avenin-9 Immunogenic Native DQ2.5 90, 18, 8 9 PYPEQQEPF 1005 ave-1 gliadin alpha avenin-9 Immunogenic Deamidated DQ2.5 90, 18, 8 9 PYPEQEEPF 1006 Avenin Av-gamma2B Immunogenic Native DQ2  8 14 QQPFVQQQQPFVQQ 1007 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVQQQQPFVQQ 1008 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVEQQQPFVQQ 1009 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVEQQQPFVQQ 1010 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVQEQQPFVQQ 1011 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVQEQQPFVQQ 1012 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVEEQQPFVQQ 1013 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVEEQQPFVQQ 1014 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVQQEQPFVQQ 1015 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVQQEQPFVQQ 1016 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVEQEQPFVQQ 1017 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVEQEQPFVQQ 1018 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVQEEQPFVQQ 1019 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVQEEQPFVQQ 1020 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 QQPFVEEEQPFVQQ 1021 Avenin Av-gamma2B in Deamidated form Immunogenic Deamidated DQ2  8 14 EQPFVEEEQPFVQQ 1022 Avenin Avenin core epitope in native form Immunogenic Native DQ2 27 9 FVQQQQQPF 1023 Avenin Avenin core epitope in Deamidated form Immunogenic Deamidated DQ2 27 9 FVQQQEQPF 1024 gamma-gliadin Glu-21 minimal epitope in considered native form Immunogenic Native DQ2 19 12 QSEQSQQPFQPQ or LMW glutenin 1025 glia-gamma 3, gamma gliadin Immunogenic Deamidated DQ2.5 90, 17, 23 9 EQPQQPYPQ glia-gamma 1b or DQ8 1026 glia-gamma 3, gamma gliadin Immunogenic Deamidated DQ2.5 90, 17, 23 9 QQPQQPYPE glia-gamma 1b or DQ8 1027 glia-gamma 3, gamma gliadin Immunogenic Deamidated DQ2.5 90, 78, 23 9 EQPEQPYPQ glia-gamma 1b or DQ8 1028 glia-gamma 3, CAUTION, 100% match to Candida protein/gamma  Immunogenic Deamidated DQ2.5 90, 78, 23 9 QQPEQPYPE glia-gamma 1b or DQ8 1029 glia-gamma 3 gamma 5 gliadin Immunogenic Deamidated DQ2.5 90, 78, 23 9 SQPEQQFPQ 1030 glia-gamma 3 gamma 5 gliadin Immunogenic Deamidated DQ2.5 90, 78, 23 9 SQPQQEFPQ 1031 glia-gamma gammaVII-gliadin Immunogenic Deamidated DQ2.5 90, 17, 25 9 QQPQQPFPE 1b, glia-gamma 4c 1032 glia-gamma c, gammaVII-gliadin Immunogenic Deamidated DQ2.5 90, 17, 25, 9 QQPEQPFPE glia-gamma 1b 8 1033 glia-gamma gammaVII-gliadin Immunogenic Deamidated DQ2.5 90, 17, 25 9 EQPEQPFPE 4c, glia-gamma 1b 1034 glia-gamma 4c gamma gliadin Immunogenic Native DQ2.5 90, 62, 78, 9 PQPQQPFCQ 81 1035 glia-gamma 4d gamma gliadin Immunogenic Deamidated DQ2.5 90, 62, 78, 9 PQPEQPFCQ 81 1036 glia-gamma 4d gamma gliadin Immunogenic Deamidated DQ2.5 90, 62, 78, 9 PQPQQPFCE 81 1037 glia-gamma 4d gamma gliadin Immunogenic Deamidated DQ2.5 90, 62, 78, 9 PQPEQPFCE 81 1038 glia-gamma 4e gliadin omega 1 Immunogenic Native DQ2.5 90, 86 9 PQPQQPFSQ 1039 glia-gamma 4e gliadin omega 1 Immunogenic Deamidated DQ2.5 90, 86 9 PQPEQPFSQ 1040 glia-omega 3 gliadin omega 3 Immunogenic Native DQ2.5 86, 90 9 PFPQPQQPI 1041 glia-omega 3 gliadin omega 3 Immunogenic Deamidated DQ2.5 86, 90 9 PFPQPEQPI 1042 glia-omega 4 CAUTION 100% match to 2 Prunus sp. peptides/ Immunogenic Native DQ2.5 86, 90 9 PQPQQPIPV gliadin omega 4 1043 glia-omega 4 gliadin omega 4 Immunogenic Deamidated DQ2.5 86, 90 9 PQPEQPIPV 1044 glia-omega 5 gliadin omega 5 Immunogenic Native DQ2.5 86, 90 9 LQPQQPFPQ 1045 glia-omega 5 gliadin omega 5 Immunogenic Deamidated DQ2.5 86, 90 9 LQPEQPFPQ 1046 Avenin Q avenin-gliadin like Immunogenic Native DQ2 or 87 14 QQPFMQQQQPFMQP DQ8 1047 Avenin Q-5 avenin-gliadin like Immunogenic Native DQ2 or 88 14 QQPFVQQQQQPFVQ DQ8 1048 glia-gamma 1 gamma gliadin 1 Immunogenic Deamidated DQ2.5 90, 74 9 PEQSFPQQQ 1049 glia-gamma 1 gamma gliadin Immunogenic Deamidated DQ2.5 90, 74 9 PEQSFPQQE 1050 ave-1 06 Avenin gliadin like Immunogenic Native DQ2.5 88 16 QYQPYPEQQQPILQ QQ 1051 ave-1 06 avenin-gliadin like Immunogenic Deamidated DQ2.5 88 16 QYQPYPEQEQPILQ QQ 1052 ave-1 04 avenin-gliadin like Immunogenic Native DQ2.5 88 16 QQYQPYPQQQPFMQ PL 1053 ave-1 04 avenin-gliadin like Immunogenic Deamidated DQ2.5 88 16 EQYQPYPEQQPFMQ PL CD1D multimer and ligand:

In one embodiment, the peptide is a human, non-human, or synthetic/engineered peptide. For example, the peptide is a Minor Histocompatibility Antigen (MiHA).

Exemplary MiHAs are known in the art, e.g., in Spierings et al. Tissue Antigens 2014 84:347-360; which is incorporated by reference herein. MiHAs are typically utilized in embodiments relating to transplantation, where it refers to epitopes that are created because of protein sequence variation in polymorphic proteins.

Residues of the antigenic peptide that engages the MHC binding groove can loosely be divided into two types: anchor residues, which engage with the MHC molecule and confer stability to the MHC-peptide complex (for example, see residues P2, P3, P5, P6, P7, and P9 in Bowness et al. 1999 Expert Reviews in Molecular Medicine 16:1-10; which is incorporated by reference herein in its entirety), and interfacial residues, which are solvent-exposed and can engage with the cognate T-cell receptor (see, e.g., residues P1, P4, and P8 in Bowness). A featureless peptide is a peptide in which anchor residues are preserved, while interfacial residues of the peptide are mutated to alanine or glycine residues to prevent TCR binding A featureless peptide-MHC is therefore a MHC peptide complex in which the presented peptide is a featureless peptide. Featureless peptide MHCs are typically used in embodiments relating to transplantation tolerance. In patients receiving MHC-mismatched solid organ or hematopoietic stem cell transplants, use of a CAL T cell presenting the relevant donor-mismatched featureless peptide-MHC CAL can permit selective depletion of recipient alloreactive T cells targeted towards this mismatched donor HLA allele.

The MHC can be a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form. In embodiments where more than one MHC unit is present, e.g., the MHC is oligomeric, the units can be provided in series, e.g., in a chain, or provided arrayed in one or more dimensions around a central point or linker, provided conjugated/bound in any geometry to a scaffold molecule, or any combination of the foregoing. Naturally occurring examples of TCR recognition domain structures in the art include MHC dimers (Lebowitz et al., 1999 Cellular Immunology 192:175-184), tetramers (Altman et al., 1996 Science 274:94-96), pentamers (proimmune.com/introduction-to-pentamers/), octamers (Guillame et al., 2003 JBC 278:4500-4509), dextramers (Batard et al., 2006 Journal of Immunological Methods 310:136-148), dodecamers (Huang et al., 2016 PNAS 113:E1890-7), lipid vesicles (Mallet-Designe et al., 2003 The Journal of Immunology 170:123-131), and quantum dots (Chattopadhyay et al., 2006 Nature Medicine 12:972-7). Each of the foregoing references is incorporated by reference herein in its entirety.

In some embodiments, the TCR recognition domain can comprise a CD1 domain (e.g., a CD1d domain), e.g., a sequence comprising an extracellular domain of CD1, (e.g., CD1). As used herein, “cluster of differentiation 1 family member d” or “CD1d” refers to a cell surface protein that displays lipid antigens to T cells. The sequences of several CD1d isoforms, and the structure of CD1d are known in the art, see, e.g., the 3 isoforms provided in the NCBI database for CD1d (Gene ID 912), and Bagchi et al., 2018 and Oleinika et al., Nature Communications 2018 9:684; which are incorporated by reference herein in their entireties. For example, isoform 1 of CD1d is SEQ ID NO: 5 (NCBI Ref Seq NP_001757.1), isoform 2 of CD1d is SEQ ID NO: 6 (NCBI Ref Seq NP_001306074.1), and isoform 3 of CD1d is SEQ ID NO: 7 (NCBI Ref Seq NP_001358690.1). In some embodiments, the CD1d domain comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to one of SEQ ID NOs: 5-7. In some embodiments, the CD1d domain comprises a sequence with at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or greater sequence identity to the extracellular domain of one of SEQ ID NOs: 5-7 (e.g., amino acids 20-301 of SEQ ID NO: 5). In some embodiments, the CD1d domain comprises a sequence with at least at least 95% sequence identity to one of SEQ ID NOs: 5-7 and retains the lipid binding activity of the wild-type reference sequence. In some embodiments, the CD1d domain comprises a sequence with at least 95% sequence identity to the extracellular domain of one of SEQ ID NOs: 5-7 (e.g., amino acids 20-301 of SEQ ID NO: 5) and retains the lipid binding activity of the wild-type reference sequence.

In some embodiments, the CD1 domain further comprises a ligand, e.g., a non-peptide ligand. Exemplary, non-limiting ligands of CD1 domains include but are not limited to:

Protein (Allele) Ligand human CD1a C-glycoside GSL-1 OCH human CD1b C-glycoside GSL-1 OCH human CD1c C-glycoside GSL-1 OCH human CD1d OCH human CD1d PBS-57

In some embodiments, a TCR recognition domain can comprise sequences or molecules in addition to the, e.g., MHC, pMHC, of CD1 domains and sequences. For example, it can further comprise other polypeptide and/or non-polypeptide components that enable multimerization. Exemplary components that permit multimerization can include biotin (non-polypeptide) and/or streptavidin polypeptide that is used to permit tetramerization.

In various embodiments, protein interaction domains are found on an extracellular portion of the respective polypeptides.

As used herein, “recognition polypeptide” refers to an extracellular polypeptide having a ligand-binding domain. In some embodiments, the ligand-binding domain can be an antibody reagent. In some embodiments, the recognition polypeptide can further comprise a protein interaction domain.

As used herein, “signaling polypeptide” refers to a transmembrane polypeptide having an intracellular signaling domain, e.g., a T cell receptor (TCR) signaling domain. In some embodiments, the signaling polypeptide can further comprise a protein interaction domain. In some embodiments, the signaling polypeptide can further comprise an extracellular protein interaction domain.

As used herein, “biomolecular interaction domain” refers to a domain that permits specific binding of two separate molecules to each other. The molecules can be or can comprise polypeptides. In some embodiments, one or both of the molecules or biomolecular interaction domains can be a non-peptide. When a pair of biomolecular interaction domains is provided herein, they permit two or more molecules to bind specifically, e.g. one of the biomolecular interaction domains can bind specifically to the second biomolecular interaction domain. In some embodiments, specific binding can occur when two separate biomolecular interaction domains, e.g., of a pair, are present. In some embodiments, specific binding can occur when three or more separate biomolecular interaction domains are present. It is noted that protein interaction domains are a type of biomolecular interaction domains and where one is specified herein, the other may always be substituted.

As used herein, when a molecule is referred to as a protein or polypeptide, it comprises a protein, peptide, or polypeptide sequence, but may comprise additional motifs, modifications, or domains of a non-proteinaceous nature. A number of exemplary biomolecular interaction domains, as well as pairs of protein interaction domains are provided elsewhere herein. In some embodiments, the biomolecular interaction domains comprise, consist, or consist essentially of proteins or polypeptides. In some embodiments, the biomolecular interaction domains comprise, consist, or consist essentially of non-proteinaceous molecules. In some embodiments, one of a pair of biomolecular interaction domains can comprise, consist, or consist essentially of proteins or polypeptides and the second of the pair of biomolecular interactions domains can comprise, consist, or consist essentially of a non-proteinaceous molecule (e.g., FITC and anti-FITC). Exemplary protein interaction domains are known in the art and can be used in embodiments of the aspects described herein.

As used herein, “protein interaction domain” refers to a domain that permits specific binding of two separate polypeptides to each other. A number of exemplary protein interaction domains, as well as pairs of protein interaction domains are provided elsewhere herein. In some embodiments, the protein interaction domains of the polypeptides of a multi-component CAL and/or CAR can bind specifically, e.g. one of the protein interaction domains can bind specifically to a second protein interaction domain of the multi-component CAL and/or CAR. In some embodiments, specific binding can occur when two separate protein interaction domains are present. In some embodiments, specific binding can occur when three or more separate protein interaction domains are present. Exemplary protein interaction domains are known in the art and can be used in embodiments of the aspects described herein.

In some embodiments of any of the aspects described herein, the protein interaction domains can be leucine zipper domains. Leucine zipper domains are a type of protein-protein interaction domain commonly found in transcription factors characterized by leucine residues evenly spaced through a α-helix. Leucine zippers may form heterodimers or homodimers. A number of leucine zipper domains, as well as their ability to bind each other, are known in the art and discussed further, e.g., in Reinke et al. JACS 2010 132:6025-31 and Thompson et al. ACS Synth Biol 2012 1:118-129; each of which is incorporated by reference herein in its entirety. In some embodiments, one leucine zipper domain is BZip (RR) and the second leucine zipper domain is AZip (EE). In some embodiments, the sequence of a BZip (RR) leucine zipper domain is MDPDLEIRAAFLRQRNTALRTEVAELEQEVQRLENEVSQYETRYGPLGGGK (SEQ ID NO: 3). In some embodiments, the sequence of a AZip (EE) leucine zipper domain is MDPDLEIEAAFLERENTALETRVAELRQRVQRLRNRVSQYRTRYGPLGGGK (SEQ ID NO: 4). Further exemplary leucine zipper domains are described in Reinke et al. JACS 2010 132:6025-31; which is incorporated by reference herein in its entirety. For example, suitable leucine zipper domains can include SYNZIP 1 to SYNZIP 48, and BATF, FOS, ATF4, ATF3, BACH1, JUND, NFE2L3, and HEPTAD. Binding affinities of various combinations of these domains are described, e.g., at FIG. 1 of Reinke et al. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 1000 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 100 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 10 nM or less. In some embodiments, a suitable pair of leucine zipper domains has a dissociation constant (Kd) of 1 nM or less.

Further exemplary pairs of protein interaction domains can include a) PSD95-D1g1-zo-1 (PDZ) domains; b) a streptavidin domain and a streptavidin binding protein (SBP) domain; and c) a PYL domain and an ABI domain.

In some embodiments of any of the aspects described herein, the protein interaction domains can be chemically-induced protein interaction domains, e.g., domains that will only bind specifically in the presence of a third molecule, e.g., a small molecule or drug. Exemplary pairs of chemically-induced protein interaction domains can include: FKBP-binding domain of mTOR (FRB) and FK506 binding protein (FKBP) (binding of which is activated by tacrolimus, everolimus, or a rapalog); cyclophilin-Fas fusion protein (CyP-Fas) and FK506 binding protein (FKBP) (binding of which is activated by FKCsA); calcineurin A (CNA) and FK506 binding protein (FKBP) (binding of which is activated by FK506); gibberellin insensitive (GIA) and gibberellin insensitive dwarf1 (GID1) (binding of which is activated by gibberellin); Snap-tag and Halo tag (binding of which is activated by HaXS); and T14-3-3-cdeltaC and C-Terminal peptides of PMA2 (CT52) (binding of which is activated by fusicoccin). Further description of chemically-induced protein interaction domains can be found in the art, e.g., Miyamoto et al. Nat Chem Biol. 2012 Mar. 25; 8(5): 465-470 and Belshaw et al. PNAS 1996 93:4604-4607; each of which is incorporated herein by reference in its entirety.

In some embodiments of any of the aspects described herein, the protein interaction domains can comprise at least one nucleotide tag and at least one zinc finger domain. Zinc finger domains are characterized by the coordination of a zinc ion in order to stabilize their tertiary structure. The particular folds that appear in zinc fingers can vary. In some embodiments, a zinc finger domain can be a nucleotide-binding zinc finger domain. In some embodiments, a zinc finger domain can be a DNA-binding zinc finger domain. In some embodiments, the protein interaction domain of the recognition polypeptide is a nucleotide tag and the extracellular protein interaction domain of the signaling polypeptide is a zinc finger domain. In some embodiments, a nucleotide tag can be a DNA tag. In some embodiments, a nucleotide tag can be a dsDNA tag comprising the entire recognition sequence for the zinc finger domain being used. Exemplary zinc finger domains and their cognate nucleotide tags are described in the art, e.g. Mali et al. Nature Methods 2013 10:403-406; which is incorporated by reference herein in its entirety. In some embodiments, a zinc finger domain can be sZF15 as described in Mali et al. Nature Methods 2013 10:403-406.

In one embodiment, the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE).

In some embodiments of any of the aspects described herein, the protein interaction domains can comprise a pair of substantially complementary nucleotide tags, e.g., fully complementary or complementary enough to hybridize specifically. The degree of complementarity necessary may vary depending on the total length of the tags and G/C content of the complementary portions. One of skill in the art can readily determine the relevant affinity necessary for tags of a given size and G/C content. In some embodiments, a nucleotide tag can be a DNA tag.

In some embodiments, a nucleotide tag can be a DNA tag. In some embodiments, a nucleotide tag can be a dsDNA tag.

In one embodiment, the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1).

In one embodiment, the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag.

In one embodiment, the protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52).

In one embodiment, the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI.

In one embodiment, the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding protein or any binding pair of biomolecular interaction domains are collectively a FITC and a FITC binding protein.

In one embodiment, the biomolecular interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of biomolecular interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein.

In some embodiments, the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain (i.e. a CAL) binds specifically to a native cell surface molecule on a NK cell, dendritic cell, or T cell. In such embodiments, a subject can be treated as described herein without administration of an engineered cell. In some embodiments, the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain (i.e. a CAL) binds specifically to a native cell surface molecule on a NK cell. In some embodiments, the biomolecular domain on the molecule comprising a TCR recognition domain and biomolecular interaction domain (i.e. a CAL) binds specifically to a native cell surface molecule on dendritic cell. Exemplary native cell surface molecules for such embodiments include, but are not limited to CD3. Other suitable cell surface molecules are the CD cell surface proteins. CDs are known in the art and one of ordinary skill can readily select one expressed by the desired cell type(s). For example, see the HCDM database at hcdm.org and the lists available on the world wide web at chemeurope.com/en/encyclopedia/List_of human_clusters_of differentiation.html; and docs.abcam.com/pdf/immunology/Guide-to-human-CD-antigens.pdf. Accordingly, exemplary biomolecular interaction domains for such embodiments include but are not limited to an anti-CD3 antibody reagent, or a Fab domain. One of skill in the art is aware of other cell surface molecules found, or found exclusively, on the cell surface of the relevant cell type(s), as well as multiple reagents that can bind (e.g., bind specifically) to such cell surface molecules.

In aspects with a single recognition polypeptide and a single signaling polypeptide that are able to bind specifically without a third polypeptide, the multiple-component CALs or CARs described herein will activate in the presence of the target ligand, thereby inducing T cell activity in the vicinity of the target ligand. Further described herein are multiple-component CALs or CARs capable of logic computation, for example, multiple-component CALs or CARs that serve as AND, OR, or NOT logic gates.

In some aspects, described herein are compositions that comprise components of a multi-component CAL and/or CAR that permits AND gate logic. In these aspects, activation of the multi-component CAL and/or CAR happens only in the presence of two target ligands; recognition of a single target ligand is not sufficient for activation. Such multi-component CALs or CARs can permit greater specificity and reduce off-target effects. Any single ligand that is a good marker for a target cell or tissue may occur elsewhere in a subject, resulting in off-target effects. However, requiring the recognition of two separate marker ligands reduces the odds of off-target activity.

In one embodiment, a nucleotide tag can be a DNA tag or dsDNA tag.

Further embodiments of AND logic gate multi-component CALs and CARs are described herein.

In some embodiments of any of the aspects described herein, the compositions comprise components of a multi-component CAL and/or CAR that are NOT logic gate. For example, recognition of a second target ligand by a second recognition polypeptide can prevent interaction (e.g. specific binding) of the signaling polypeptide and first recognition polypeptide. Such embodiments can permit suppression of T cell activity in inappropriate and/or off-target tissues. For example, the second target ligand can be a marker of a tissue that is particularly sensitive to T cell activity, is a known area of off-target activity, and/or shares markers with the desired target tissue. In some embodiments, in a NOT gate multi-component CAL and/or CAR, the second target ligand is not a ligand found in the target tissue and/or cells, e.g., in or on a disease T cell. In some embodiments, the second target ligand of a NOT logic gate multi-component CAL and/or CAR is found on a healthy and/or non-target cell and not on a diseased and/or target cell. Various 2- and 3-dimensional configurations of such pairs of nucleotide pairs are known in the art.

In some embodiments, the target ligand recognized by the second recognition polypeptide is found on a healthy and/or non-target cell and not on a diseased and/or target cell. In some embodiments, the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the first recognition polypeptide have a greater affinity than the protein interaction domain of the signaling polypeptide and the protein interaction domain of the first recognition polypeptide. In some embodiments, the protein interaction domain of the second recognition polypeptide and the protein interaction domain of the signaling polypeptide have a greater affinity than the protein interaction domain of the signaling polypeptide and the protein interaction domain of the first recognition polypeptide. Relative binding affinities can be determined experimentally, e.g., by binding affinity assays known in the art and relative binding affinities are known for a number of combinations of protein interaction domains described herein, see, e.g. Reinke et al. JACS 2010 132:6025-31; which is incorporated by reference herein in its entirety. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domains can be at least 2× greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domains can be at least 5× greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain. In some embodiments, the binding affinity of the recognition polypeptide protein interaction domains can be at least 10× greater than the binding affinity of the first recognition polypeptide protein interaction domain and the signaling polypeptide interaction domain.

As used herein, “target ligand” refers to a molecule in or on a cell which can be bound by a ligand-binding domain. Non-limiting examples of such molecules can include polypeptides, lipids, saccharides, and the like. In some embodiments, the target ligand can be an extracellular molecule. In some embodiments, the target ligand can be a cell surface molecule.

In some embodiments, e.g., those relating to a multi-component CAL and/or CAR with a single recognition polypeptide or an AND gate multi-component CAL and/or CAR, the target ligand (e.g. the first and/or second target ligand) can be a ligand expressed in a target tissue. In some embodiments, the target ligand can be expressed constitutively in the target tissue and/or cell. In some embodiments, the target ligand can be expressed exclusively in the target tissue and/or cell. In some embodiments, the target ligand can be expressed at a higher level in the target tissue and/or cell than in other tissues and/or cells. As recognition of a target ligand in embodiments relating to a multi-component CAL and/or CAR with a single recognition polypeptide or an AND gate multi-component CAL and/or CAR can result in T cell activation (e.g. cell killing activity of the cell comprising the target ligand), the target ligand can be selected to target T cell activity in a desirable and/or therapeutic way, e.g., by targeting a disease cell. In some embodiments, a target ligand is a ligand found in/on a diseased and/or target cell. In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is a ligand found in/on a diseased and/or target cell. In some embodiments, a target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is a ligand found on a diseased and/or target cell and not on a healthy and/or non-target cell. In some embodiments, the diseased cell is an autoreactive or alloreactive T cell. In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR is found on the surface of a disease cell. In some embodiments, a recognition polypeptide that can specifically bind with a signaling polypeptide or is a portion of an AND gate multi-component CAL and/or CAR specifically binds to a target ligand on the surface of a disease cell, e.g. as compared to binding to normal cells.

In some embodiments, a composition and/or cell described herein can further comprise a second multi-component CAL and/or CAR according to any of the aspects and embodiment described herein for the first multi-component CAL and/or CAR. By way of non-limiting example, a second CAL and/or CAR can be designed to bind specifically to (and, e.g., be activated by or inhibited by) different target ligands than those to which the first multi-component CAL and/or CAR specifically binds (and, e.g. is activated by or inhibited by). This can provide increased specificity, reduced off-target effects, and/or reduced effective dosages for the methods described herein. In some embodiments, the recognition domain of second multi-component CAL and/or CAR bind specifically to different target ligands than those bound by the recognition domain of the first multi-component CAL and/or CAR. In some embodiments, the antibody reagents of second multi-component CAL and/or CAR bind specifically to different target ligands than those bound by the antibody reagents of the first multi-component CAL and/or CAR.

In some embodiments, the second multi-component CAL and/or CAR can comprise an inhibitory intracellular signaling domain, e.g., T cell receptor (TCR) signaling domain, e.g., one that inhibits engineered cell, e.g., T cell, activity. In such embodiments, the second multi-component can therefore be designed to operate in opposition to the first multi-component CAL and/or CAR, e.g. permitting inhibition of T cell activation while the first multi-component CAL and/or CAR permits activation of T cell activity. Inhibitory intracellular TCR signaling domains are known in the art and can include, by way of non-limiting example, PD1; CTLA4; BTLA; KIR; LAG-3; TIM-3; A2aR; LAIR-1; and TGIT. In one embodiment, non-active mimetics of an activating TCR can be used.

In some embodiments, described herein is a composition comprising a cell, a CAL, and a CAR (e.g., with the CAR comprising either an inhibitory domain or costimulation/activation domain).

In one embodiment, the TCR recognition domain comprises a MHC allogeneic to the cell. In one embodiment, the TCR recognition domain comprises a peptide allogeneic to the cell. In one embodiment, the TCR recognition domain comprises a MHC allogeneic to the target cell. In one embodiment, the TCR recognition domain comprises a peptide allogeneic to the target cell. In one embodiment, the TCR recognition domain comprises a non-self peptide relative to the target cell.

In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with the signaling polypeptide of the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain is a ligand found on a healthy and/or non-target cell. In some embodiments, the target ligand specifically bound by a recognition polypeptide that can specifically bind with the signaling polypeptide of the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain is a ligand found on a healthy and/or non-target cell and not on a diseased and/or target cell. In some embodiments, the second multi-component CAL and/or CAR comprising an inhibitory intracellular signaling domain can be an OR logic gate according to any of the embodiments described herein and the second target ligand can be a ligand found in/on, or specific to, diseased cells.

In some embodiments of any of the aspects, a ligand-binding domain can comprise or consist essentially of an antibody reagent. In some embodiments, the antibody reagent can be an immunoglobulin molecule, a monoclonal antibody, a chimeric antibody, a CDR-grafted antibody, a human antibody, a humanized antibody, a Fab, a Fab′, a F(ab′)2, a Fv, a disulfide linked Fv, a scFv, a single domain antibody, a diabody, a multispecific antibody, a dual specific antibody, an anti-idiotypic antibody, and/or a bispecific antibody.

In some embodiments, the intracellular signaling domain can be a T-cell activation domain. In some embodiments, the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCRζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD3ζ, CD22, CD79a, CD79b, CD66d, CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, TRIM, and ZAP70. In some embodiments, the signaling domain can be a paralog or ortholog of any of the foregoing.

Multi-Component CAR/CAL Cells

Presented herein are cells that express the compositions or multi-component CARs and/or CALs presented herein. A cell can be any cell, for example, any mammalian cell, e.g., a human cell. In one embodiment, the cell is a dendritic cell, regulatory T cell, or effector T cell. In one embodiment the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.) (CAL-T); regulatory T cell, effector T cell, natural killer cell (CAL NK), or any other myeloid cell.

In one aspect, described herein is an engineered cell expressing and/or comprising one or more multi-component CARs/CALs, or a composition comprising the same as described herein, e.g., at least one signaling polypeptide and at least one recognition polypeptide. In some embodiments, the cell is a natural killer (NK) cell, dendritic cell, regulatory T cell, effector T cell. Such cells expressing and/or comprising both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAR/CAL are referred to herein as “complete multi-component CAR/CAL” cells. In some embodiments, a complete multi-component CAR/CAL cell expresses both a signaling polypeptide (e.g., a CAR) and at least one recognition polypeptide (e.g. a CAL or adaptor as described elsewhere herein) of a multi-component CAL and/or CAR. In some embodiments, a complete multi-component CAL and/or CAR cell comprises nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR. In some embodiments, a signaling polypeptide is present on the membrane of a cell. In some embodiments, the one or more recognition polypeptides are present in the extracellular space, e.g., the recognition polypeptide(s) can be expressed and secreted by the cell or the cell can be contacted by recognition polypeptides provided from another source (e.g. produced synthetically or by another cell and optionally, purified or processed before the contacting step).

In any of the aspects described herein, e.g., those relating to either a complete or partial multi-component CAL and/or CAR cell, the recognition and/or signaling polypeptide can be under the control of an inducible and/or repressible promoter. Such promoters allow the expression of the polypeptide to be increased or decreased as desired and are in contrast to constitutive promoters. The term “constitutively active promoter” refers to a promoter of a gene which is expressed at all times within a given cell. Exemplary promoters for use in mammalian cells include cytomegalovirus (CMV), Elongation Factor 1a (EF1a), and the like. The term “inducible promoter” refers to a promoter of a gene which can be expressed in response to a given signal, for example addition or reduction of an agent. Non-limiting examples of an inducible promoter are promoters that are regulated in a specific tissue type, a promoter regulated by a steroid hormone, by a polypeptide hormone (e.g., by means of a signal transduction pathway), or by a heterologous polypeptide (e.g., the tetracycline-inducible systems, “Tet-On” and “Tet-Off”; see, e.g., Clontech Inc., CA, Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89:5547, 1992, and Paillard, Human Gene Therapy 9:983, 1989; each of which are incorporated by reference herein in its entirety). In some embodiments, expression of the polypeptide can be precisely regulated, for example, by using an inducible regulatory sequence that is sensitive to certain physiological regulators, e.g., circulating glucose levels, or hormones (Docherty et al., 1994, FASEB J. 8:20-24). Such inducible expression systems, suitable for the control of expression in cells or in mammals include, for example, regulation by ecdysone, by estrogen, progesterone, tetracycline, chemical inducers of dimerization, and isopropyl-beta-D1-thiogalactopyranoside (IPTG). A person skilled in the art would be able to choose the appropriate regulatory/promoter sequence based on the intended use of the polypeptide.

In some embodiments, the expression of one or more of the recognition or signaling polypeptides can be constitutive. In some embodiments, the expression of one or more of the recognition or signaling polypeptides can be transient. Transient expression can be achieved by, e.g., use of transient and/or inducible expression promoters or by use of transient vectors, e.g. those that do not incorporate into the genome and/or persist in the target cell. By way of non-limiting example, derivatives of viruses such as the bovine papillomavirus (BPV-1), or Epstein-Barr virus (pHEBo, pREP-derived and p205) can be used for transient expression of nucleic acids in eukaryotic cells. For other suitable expression systems as well as general recombinant procedures, see Molecular Cloning A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch and Maniatis (Cold Spring Harbor Laboratory Press: 1989) Chapters 16 and 17; which is incorporated by reference herein in its entirety. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the recognition polypeptide can be transiently expressed. In some embodiments, the recognition polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the signaling polypeptide can be transiently expressed. In some embodiments, the recognition polypeptide of a multi-component CAL can be constitutively expressed and the signaling polypeptide can be provided exogenously. In some embodiments, the recognition polypeptide of a multi-component CAL can be transiently expressed and the signaling polypeptide can be provided exogenously. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be constitutively expressed and the recognition polypeptide can be provided exogenously. In some embodiments, the signaling polypeptide of a multi-component CAL and/or CAR can be transiently expressed and the recognition polypeptide can be provided exogenously.

The CARs and CALs described herein can be produced according to any method known in the art, e.g., recombinant expression or peptide synthesis. Exemplary methods can include, the NIH Tetramer Core Facility's MHC expression protocols (available on the world wide web at tetramer.yerkes.emory.edu/support/protocols#1); ProImmune's pentamer protocols (available on the world wide web at proimmune.com/protocols-2/); Immudex's dextramer protocols (available on the word wide web at immudex.com/resources/protocols/) and the CAR T cell production protocols provided in the “Primary Human T cells Isolation and Culture” and “Lentiviral Transduction of Human T cells” sub-sections in “Method Details” section of Cho et al., 2018 Cell 173:1426-1438; each of which is incorporated by reference herein in its entirety.

In one aspect, described herein is a method of killing a target cell, the method comprising contacting the cell with a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein. In some embodiments, the target cell can be a diseased cell, e.g., an autoreactive or alloreactive T cell. In one aspect, described herein is a method of treating or preventing a disease, e.g., an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection, or GvHD, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein. In one aspect, described herein is a method of treating or preventing autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein.

Another aspect provided herein is a method of preventing and/or treating a malignant T cell condition in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein. In one aspect, described herein is a method of treating or preventing a malignant T cell condition in a subject, comprising administering a complete multi-component CAR/CAL, CAR and/or CAL as according to any of the embodiments described herein. In one aspect, described herein is a method of treating or preventing a malignant T cell condition in a subject, comprising administering a complete multi-component CAR cell, CAR, and/or CAL according to any of the embodiments described herein

In some embodiments, the complete multi-component CAL and/or CAR cell can be autologous or allogeneic to the subject. In some embodiments, the complete multi-component CAL and/or CAR cell can be derived and/or descended from a cell obtained from the subject or a third party and has been modified ex vivo to comprise the at least one multi-component CAL and/or CAR, e.g., genetically engineered to comprise nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR. In some embodiments, the method can further comprise the steps of obtaining a cell from a subject (e.g. a dendritic cell, regulatory T cell, or effector T cell), altering the cell to comprise nucleic acid sequences encoding both a signaling polypeptide and at least one recognition polypeptide of a multi-component CAL and/or CAR, and then administering the cell to the subject.

In one embodiment, the engineered cell is further modified to lack or have reduced expression of the native MHCI/II, e.g., as measured on the cell surface. Methods for engineering a cell to reduce or eliminate the native MHCI/II are known in the art and include, e.g., expression of RNA interference (such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.), expression of an inhibitory oligonucleotide, nuclease-based inhibition (such as CRISPR, TALEN, Meganuclease, etc.), and expression of a KDEL-motif (SEQ ID NO: 2749) containing binding protein capable of restricting MHCI/II to the endoplasmic reticulum. One skilled in the art can assess whether knockdown of native MHCI/II is achieved by assessing mRNA or protein levels of MHCI/II via, e.g., PCR-based assays or western blotting, respectively.

In one embodiment, the cell is further engineered to knockout the native MHCI/II. Methods for engineering a cell to knockout the native MHCI/II are known in the art and include, e.g., expression of RNA interference (such as short hairpin RNA, small interfering RNA, double stranded RNA, etc.), expression of an inhibitory oligonucleotide, nuclease-based inhibition (such as CRISPR, TALEN, Meganuclease, etc.). One skilled in the art can assess whether knockout of native MHCI/II is achieved by assessing mRNA or protein levels of MHCI/II via, e.g., PCR-based assays or western blotting, respectively.

In one aspect, described herein is an engineered cell expressing and/or comprising one or more of the compositions according to any of the embodiments described herein. In one aspect, described herein is an engineered cell expressing and/or comprising one or more multi-component CAL and/or CAR signaling polypeptides according to any of the embodiments described herein. In some embodiments, the cell is a dendritic cell, regulatory T cell, or effector T cell. In some embodiments, the cell is a T cell. Such cells expressing and/or comprising a multi-component CAL and/or CAR signaling polypeptide are referred to herein as “partial multi-component CAL” cells or “partial multi-component CAR” cells. In some embodiments, the partial multi-component CAL and/or CAR cell does not express, e.g., does not comprise a nucleic acid sequence encoding, a multi-component CAL and/or CAR recognition polypeptide. In some embodiments, a partial multi-component CAL and/or CAR cell comprises a nucleic acid sequence encoding at least one multi-component CAL and/or CAR signaling polypeptide. In some embodiments, the multi-component CAL and/or CAR signaling polypeptide is present on the membrane of the cell, e.g., is expressed as a transmembrane protein at detectable levels. In some embodiments, the signaling polypeptide further comprises a secondary protein interaction domain that specifically binds with the protein interaction domain of the second recognition polypeptide, e.g., the signaling polypeptide is part of an AND gate multi-component CAL and/or CAR as described elsewhere herein. In some embodiments, the cell can further comprise a second multi-component CAL and/or CAR signaling polypeptide, e.g., a signaling polypeptide that is part of a second multi-component CAL and/or CAR according to any of the embodiments described herein.

In one aspect, described herein is a method of killing a target cell, the method comprising contacting the target cell with a partial multi-component CAL and/or CAR cell according to any of the embodiments described herein and contacting the target cell with at least one recognition polypeptide of the multi-component CAL and/or CAR. In some embodiments, the target cell can be a diseased cell, e.g., an autoreactive or alloreactive T cell. In some embodiments, the target cell can be a diseased cell, e.g., a cancer cell (e.g., a T cell- or T cell precursor cell-derived neoplasm, hereafter referred to as “T cell neoplasms”. In some embodiments, the method can further comprise the steps of obtaining a cell from a subject (e.g. a NK cell, a dendritic cell, regulatory T cell, or effector T cell), altering the cell to comprise a nucleic acid sequence encoding a signaling polypeptide of a multi-component CAL and/or CAR, and then administering the cell to the subject.

In some embodiments of any of the methods described herein, a pair of protein interaction domains of a multi-component CAL and/or CAR can comprise chemically induced binding domains and the method can further comprise administering a compound that induces binding of the domains. In some embodiments, when one protein interaction domain is FKBP-binding domain of mTOR (FRB) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering tacrolimus, a rapalog, or everolimus. In some embodiments, when one protein interaction domain is cyclophilin-Fas fusion protein (CyP-Fas) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering FKCsA. In some embodiments, when one protein interaction domain is calcineurin A (CNA) and a second protein interaction domain is FK506 binding protein (FKBP), the method further comprises administering FK506. In some embodiments, when one protein interaction domain is gibberellin insensitive (GIA) and a second protein interaction domain is gibberellin insensitive dwarf1 (GID1), the method further comprises administering gibberellin. In some embodiments, when one protein interaction domain is Snap-tag and a second protein interaction domain is Halo tag, the method further comprises administering HaXS. In some embodiments, when one protein interaction domain is T14-3-3-cdeltaC and a second protein interaction domain is C-Terminal peptides of PMA2 (CT52), the method further comprises administering fusicoccin.

In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be engineered. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be transgenic. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be recombinant. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be heterologous to a human T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a T cell. In some embodiments of any of the aspects described herein, a recognition and/or signaling polypeptide of a multi-component CAL and/or CAR can be exogenous to a human T cell.

It is specifically contemplated herein that each of the individual embodiments described herein can be combined, e.g., in a single cell. By way of non-limiting example, a single cell could comprise a first complete multi-component CAL and/or CAR and a second partial multi-component CAL and/or CAR, wherein each multi-component CAL and/or CAR can be according to any of the embodiments described herein.

In some embodiments, the methods described herein relate to CAL-immune cell therapies. In some embodiments, the methods described herein relate to CAR-immune cell therapies such as CAR-T therapy. Standard CAR-T and related therapies relate to adoptive cell transfer of immune cells (e.g. T cells) expressing a CAR that binds specifically to a targeted cell type (e.g. disease cells, e.g., autoreactive or alloreactive T cells) to treat a subject, e.g., for an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD.

In some embodiments, the cells administered as part of the therapy can be autologous to the subject. In some embodiments, the cells administered as part of the therapy are not autologous to the subject. In some embodiments, the cells are engineered and/or genetically modified to express a multi-component CAL and/or CAR, or portion thereof as described herein. Further discussion of CAR-T therapies can be found, e.g., in Maus et al. Blood 2014 123:2624-35; Reardon et al. Neuro-Oncology 2014 16:1441-1458; Hoyos et al. Haematologica 2012 97:1622; Byrd et al. J Clin Oncol 2014 32:3039-47; Maher et al. Cancer Res 2009 69:4559-4562; and Tamada et al. Clin Cancer Res 2012 18:6436-6445; each of which is incorporated by reference herein in its entirety.

In some embodiments, the technology described herein relates to a syringe or catheter, including an organ-specific catheter (e.g., renal catheter, biliary catheter, cardiac catheter, etc.), comprising a therapeutically effective amount of a composition described herein.

Methods of Treatment

In some embodiments, the methods described herein relate to the treatment or prevention of transplant rejection in a subject having a cell, tissue, or organ transplant with one or more compositions, CALs, CARs, or cells as described herein. In another embodiment, the methods described herein relate to the treatment or prevention of GvHD in a subject having a cell, tissue, or organ transplant with one or more compositions, CALs, CARs, or cells as described herein.

As used herein, “GvHD” refers to a disease characterized by the active process of donor cells attacking the recipient's own cells. GvHD can develop soon after a transplant, e.g., within weeks or months (acute GvHD), or can occur much later after the transplant, e.g., at least 3-6 months later (chronic GvHD). Symptoms of acute GvHD include, but are not limited to, skin rash or blisters, abdominal pain or discomfort, diarrhea, jaundice, and edema. Symptoms of chronic GvHD include, but are not limited to, changes to skin or nail texture, hair loss or thinning, muscle pain or weakness, blurred vision, mouth sores, shortness of breath, persistent cough, abdominal pain or discomfort, and diarrhea.

A subject can be identified as having or be at risk of having GvHD by a skilled clinician. Diagnostic tests useful in identifying a subject having GvHD are known in the art and will vary based on the type of transplant the subject has received. The diagnosis of GvHD is made by, for example, physical examination for the signs and symptoms for GvHD known in the art, serologic testing for dysfunction of the liver, gall bladder, kidney, and hematopoietic cells, histologic analysis of biopsies obtained from affected organs, and radiologic imaging of affected organs. In one embodiment, the method further comprises administering at least a second therapeutic. In one embodiment, the composition, CARs, CALs, or cells described herein are administered in combination with Abatacept (Orencia®) or Belatacept (Nulojix®). Abatacept and Belatacept, developed by Bristol-Meyers Squibb, are fusion proteins composed of the Fc region of the immunoglobulin IgG1 fused to the extracellular domain of CTLA-4. Abatacept is currently approved by the FDA for treatment of rheumatoid arthritis. Belatacept, which only differs from Abatacept by two amino acids, is an immunosuppressant intended to prevent rejection following a kidney transplant.

In one embodiment, the transplant is vascularized composite allograft (VCA). In one embodiment, the transplant is any human or non-human cell, tissue, or organ. In another embodiment, the transplant is any type of transplants procedures, e.g., any heart transplant, any lung transplant, any liver transplant, any pancreas transplant, any cornea transplant, any trachea transplant, any kidney transplant, any skin transplant, any pancreatic islet cell transplant, any allograft (e.g., a transplantation of allogeneic tissue), any xenograft (e.g., a transplantation of xenogeneic tissue), or any autograft (e.g., a transplantation of tissue). A skilled practitioner will be able to perform a transplant or identify a subject having had a transplant using standard procedural protocols.

In some embodiments, the methods described herein relate to the treatment or prevention of an autoimmune diseases or conditions, or hypersensitivity reaction I-IV, or immune reaction against foreign therapeutic proteins/molecules, or T cell mediated inflammation or immune response; with compositions, CARs, CALs, or cells as described herein. Subjects having an autoimmune disease can be identified by a physician using current methods of diagnosing an autoimmune disease. Symptoms and/or complications of an autoimmune disease which characterize these conditions and aid in diagnosis are well known in the art and include but are not limited to, fatigue, achy muscles, swelling and redness, low-grade fever, numbness or tingling of the hands or feet, hair loss, and/or skin rash. Tests that may aid in a diagnosis of, e.g. autoimmune disease include, but are not limited to, blood counts, and an antinuclear antibody test (ANA). A family history of autoimmune disease, or having risk factors for autoimmune disease (e.g. gender, age, ethnicity, and exposure to environmental agents, such as procainamide, hydralazine, mercury, gold, or silver) can also aid in determining if a subject is likely to have autoimmune disease or in making a diagnosis of autoimmune disease.

As used herein, the term “autoimmune disease”, “autoimmune condition”, or “autoimmune disease or disorder” herein is a disease or disorder arising from and directed against an individual's own tissues or a co-segregate or manifestation thereof or resulting condition therefrom.

Auto-immune related diseases and disorders arise from an overactive and/or abnormal immune response of the body against substances (autoantigens) and tissues normally present in the body, otherwise known as self or autologous substance. This dysregulated inflammatory reaction causes an exaggerated response by macrophages, granulocytes, lymphocytes, and/or T-lymphocytes leading to abnormal tissue damage and cell death. Subsequent loss of function is associated with inflammatory tissue damage.

Autoantigens, as used herein, are endogenous proteins or fragments thereof that are involved in or elicit this pathogenic immune response. Autoantigen can be any substance, or a portion thereof normally found within a mammal that, in an autoimmune disease, becomes the primary (or a primary, or secondary) target of attack by the immune system. The term also includes antigenic substances that induce conditions having the characteristics of an autoimmune disease when administered to mammals. Additionally, the term includes peptidic subclasses consisting essentially of immunodominant epitopes or immunodominant epitope regions of autoantigens. Immunodominant epitopes or regions in induced autoimmune conditions are fragments of an autoantigen that can be used instead of the entire autoantigen to induce the disease. In humans afflicted with an autoimmune disease, immunodominant epitopes or regions are fragments of antigens specific to the tissue or organ under autoimmune attack and recognized by a substantial percentage (e.g. a majority though not necessarily an absolute majority) of autoimmune attack T-cells.

Autoantigens that are known to be associated with autoimmune disease include myelin proteins with demyelinating diseases, e.g. multiple sclerosis and experimental autoimmune myelitis; collagens and rheumatoid arthritis; insulin, proinsulin, glutamic acid decarboxylase 65 (GAD65); and islet cell antigen (ICA512; ICA12) for insulin dependent diabetes.

A common feature in a number of autoimmune related diseases and inflammatory conditions is the involvement of pro-inflammatory CD4+ T cells. These T cells are responsible for the release of inflammatory, Th1 type cytokines. Cytokines characterized as Th1 type include interleukin 2 (IL-2), γ-interferon, TNFα and IL-12. In some embodiments, cytokines characterized as Th1 type include interleukin 2 (IL-2), interferon γ, and TNFα. Such pro-inflammatory cytokines act to stimulate the immune response, in many cases resulting in the destruction of autologous tissue. Cytokines associated with suppression of T cell response are the Th2 type, and include IL-10, IL-4 and TGF-β. It has been found that Th1 and Th2 type T cells may use the identical antigen receptor in response to an immunogen; in the former producing a stimulatory response and, in the latter, a suppressive response.

T cell mediated inflammation, or a T cell mediated immune response is inflammation and/or an immune response in which T cells and/or T cell activity contributes to or originates the inflammation/immune response.

As used herein, “inflammation” refers to the complex biological response to harmful stimuli, such as pathogens, damaged cells, or irritants. Inflammation is a protective attempt by the organism to remove the injurious stimuli as well as initiate the healing process for the tissue. Accordingly, the term “inflammation” includes any cellular process that leads to the production of pro-inflammatory cytokines, inflammation mediators and/or the related downstream cellular events resulting from the actions of the cytokines thus produced, for example, fever, fluid accumulation, swelling, abscess formation, and cell death. Inflammation can include both acute responses (i.e., responses in which the inflammatory processes are active) and chronic responses (i.e., responses marked by slow progression and formation of new connective tissue). Acute and chronic inflammation may be distinguished by the cell types involved. Acute inflammation often involves polymorphonuclear neutrophils; whereas chronic inflammation is normally characterized by a lymphohistiocytic and/or granulomatous response. An inflammatory condition is any disease state characterized by inflammatory tissues (for example, infiltrates of leukocytes such as lymphocytes, neutrophils, macrophages, eosinophils, mast cells, basophils and dendritic cells) or inflammatory processes which provoke or contribute to the abnormal clinical and histological characteristics of the disease state.

As used herein, an “immune response” refers to a response by a cell of the immune system, such as a B cell, T cell (CD4 or CD8), regulatory T cell, antigen-presenting cell, dendritic cell, monocyte, macrophage, NKT cell, NK cell, basophil, eosinophil, or neutrophil, to a stimulus (e.g., to an a disease, an antigen, or healthy cells, e.g., in the case of autoimmunity). In some embodiments of the aspects described herein, an immune response is a T cell response, such as a CD4+ response or a CD8+ response. Such responses by these cells can include, for example, cytotoxicity, proliferation, cytokine or chemokine production, trafficking, or phagocytosis, and can be dependent on the nature of the immune cell undergoing the response. Stimulation of an immune response refers to an induction or increase of the immune response. Suppression of an immune response refers to an elimination or decrease of the immune response.

A “cell-mediated immune response” is one mediated by T-cells and/or other white blood cells. A “cell-mediated immune response” is elicited by the presentation of antigenic epitopes in association with Class I or Class II molecules of the major histocompatibility complex (MHC), CD1 or other non-classical MHC-like molecules. This activates antigen-specific CD4+ T helper cells or CD8+ cytotoxic lymphocyte cells (“CTLs”). CTLs have specificity for peptide antigens that are presented in association with proteins encoded by classical or non-classical MI-ICs and expressed on the surfaces of cells. CTLs help induce and promote the intracellular destruction of intracellular microbes, or the lysis of cells infected with such microbes. Another aspect of cellular immunity involves an antigen-specific response by helper T-cells. Helper T-cells act to help stimulate the function, and focus the activity of, nonspecific effector cells against cells displaying peptide or other antigens in association with classical or non-classical MHC molecules on their surface. A “cell-mediated immune response” also refers to the production of cytokines, chemokines and other such molecules produced by activated T-cells and/or other white blood cells, including those derived from CD4+ and CD8+ T-cells. The stimulation of a cell-mediated immunological response may be determined by a number of assays, such as by lymphoproliferation (lymphocyte activation) assays, CTL cytotoxic cell assays, by assaying for T-lymphocytes specific for the antigen in a sensitized subject, or by measurement of cytokine production by T cells in response to re-stimulation with antigen. Such assays are well known in the art. See, e.g., Erickson et al. (1993) J. Immunol. 151:4189-4199; and Doe et al. (1994) Eur. J. Immunol. 24:2369-2376.

In some embodiments, the T cell mediated immune response is a response to a drug administered to the subject. It is contemplated herein that the present technology can be utilized for depletion of anti-drug specific T cells to prevent immune responses against administered biologics, cell therapies, and/or gene therapies. For example, the methods and compositions described herein can be used to prevent or treat anti-AAV and anti-transgene immune responses for administered adeno-associated virus (AAV) gene therapies, preventing or treating immune to responses to genome editing agents such as CRISPR/Cas9, Transcription activator-like effector nucleases (TALENs), or Zinc Finger Nucleases (ZFNs), and preventing or treating immune responses to enzyme replacement therapies such as recombinant human acid α-glucosidase (Pompe disease), α-L-iduronidase (Mucopolysaccharidosis I), and α-galactosidase (Fabry disease).

In one embodiment of any one of the methods described, the autoimmune disorder is selected from the group consisting of thyroiditis, type 1 diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome, Addison's disease, and Raynaud's phenomenon, Goodpasture's disease, arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gout or gouty arthritis, acute gouty arthritis, acute immunological arthritis, chronic inflammatory arthritis, degenerative arthritis, type II collagen-induced arthritis, infectious arthritis, Lyme arthritis (e.g., post treatment Lyme disease syndrome), proliferative arthritis, psoriatic arthritis, Still's disease, vertebral arthritis, and juvenile-onset rheumatoid arthritis, arthritis chronica progrediente, arthritis deformans, polyarthritis chronica primaria, reactive arthritis, and ankylosing spondylitis), palindromic arthritis, inflammatory hyperproliferative skin diseases, psoriasis such as plaque psoriasis, guttate psoriasis, pustular psoriasis, and psoriasis of the nails, atopy including atopic diseases such as hay fever and Job's syndrome, dermatitis including contact dermatitis, chronic contact dermatitis, exfoliative dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, nummular dermatitis, seborrheic dermatitis, non-specific dermatitis, primary irritant contact dermatitis, and atopic dermatitis, x-linked hyper IgM syndrome, allergic intraocular inflammatory diseases, urticaria such as chronic allergic urticaria and chronic idiopathic urticaria, including chronic autoimmune urticaria, myositis, polymyositis/dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic scleroderma), sclerosis such as systemic sclerosis, multiple sclerosis (MS) such as spino-optical MS, primary progressive MS (PPMS), and relapsing remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriosclerosis, sclerosis disseminata, ataxic sclerosis, neuromyelitis optica (NMO), inflammatory bowel disease (IBD) (for example, Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis such as ulcerative colitis, colitis ulcerosa, microscopic colitis, collagenous colitis, colitis polyposa, necrotizing enterocolitis, and transmural colitis, and autoimmune inflammatory bowel disease), bowel inflammation, pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, respiratory distress syndrome, including adult or acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, an autoimmune hematological disorder, rheumatoid spondylitis, rheumatoid synovitis, hereditary angioedema, cranial nerve damage as in meningitis, herpes gestationis, pemphigoid gestationis, pruritis scroti, autoimmune premature ovarian failure, sudden hearing loss due to an autoimmune condition, IgE-mediated diseases such as anaphylaxis and allergic and atopic rhinitis, encephalitis such as Rasmussen's encephalitis and limbic and/or brainstem encephalitis, uveitis, such as anterior uveitis, acute anterior uveitis, granulomatous uveitis, nongranulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome such as chronic or acute glomerulonephritis such as primary GN, immune-mediated GN, membranous GN (membranous nephropathy), idiopathic membranous GN or idiopathic membranous nephropathy, membrano- or membranous proliferative GN (MPGN), including Type I and Type II, and rapidly progressive GN, proliferative nephritis, autoimmune polyglandular endocrine failure, balanitis including balanitis circumscripta plasmacellularis, balanoposthitis, erythema annulare centrifugum, erythema dyschromicum perstans, eythema multiform, granuloma annulare, lichen nitidus, lichen sclerosus et atrophicus, lichen simplex chronicus, lichen spinulosus, lichen planus, lamellar ichthyosis, epidermolytic hyperkeratosis, premalignant keratosis, pyoderma gangrenosum, allergic conditions and responses, allergic reaction, eczema including allergic or atopic eczema, asteatotic eczema, dyshidrotic eczema, and vesicular palmoplantar eczema, asthma such as asthma bronchiale, bronchial asthma, and auto-immune asthma, conditions involving infiltration of T cells and chronic inflammatory responses, immune reactions against foreign antigens such as fetal A-B-O blood groups during pregnancy, chronic pulmonary inflammatory disease, autoimmune myocarditis, leukocyte adhesion deficiency, lupus, including lupus nephritis, lupus cerebritis, pediatric lupus, non-renal lupus, extra-renal lupus, discoid lupus and discoid lupus erythematosus, alopecia lupus, systemic lupus erythematosus (SLE) such as cutaneous SLE or subacute cutaneous SLE, neonatal lupus syndrome (NLE), and lupus erythematosus disseminatus, juvenile onset (Type I) diabetes mellitus, including pediatric insulin-dependent diabetes mellitus (IDDM), adult onset diabetes mellitus (Type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, diabetic retinopathy, diabetic nephropathy, diabetic large-artery disorder, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, sarcoidosis, granulomatosis including lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitides, including vasculitis, large-vessel vasculitis (including polymyalgia rheumatica and giant-cell (Takayasu's) arteritis), medium-vessel vasculitis (including Kawasaki's disease and polyarteritis nodosa/periarteritis nodosa), microscopic polyarteritis, immunovasculitis, CNS vasculitis, cutaneous vasculitis, hypersensitivity vasculitis, necrotizing vasculitis such as systemic necrotizing vasculitis, and ANCA-associated vasculitis, such as Churg-Strauss vasculitis or syndrome (CSS) and ANCA-associated small-vessel vasculitis, temporal arteritis, autoimmune aplastic anemia, Coombs positive anemia, Diamond Blackfan anemia, hemolytic anemia or immune hemolytic anemia including autoimmune hemolytic anemia (AIHA), pernicious anemia (anemia perniciosa), Addison's disease, pure red cell anemia or aplasia (PRCA), Factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases involving leukocyte diapedesis, CNS inflammatory disorders, multiple organ injury syndrome such as those secondary to septicemia, trauma or hemorrhage, antigen-antibody complex-mediated diseases, anti-glomerular basement membrane disease, anti-phospholipid antibody syndrome, allergic neuritis, Behcet's disease/syndrome, Castleman's syndrome, Goodpasture's syndrome, Reynaud's syndrome, Sjogren's syndrome, Stevens-Johnson syndrome, pemphigoid such as pemphigoid bullous and skin pemphigoid, pemphigus (including pemphigus vulgaris, pemphigus foliaceus, pemphigus mucus-membrane pemphigoid, and pemphigus erythematosus), autoimmune polyendocrinopathies, Reiter's disease or syndrome, an immune complex disorder such as immune complex nephritis, antibody-mediated nephritis, polyneuropathies, chronic neuropathy such as IgM polyneuropathies or IgM-mediated neuropathy, and autoimmune or immune-mediated thrombocytopenia such as idiopathic thrombocytopenic purpura (ITP) including chronic or acute ITP, scleritis such as idiopathic cerato-scleritis, episcleritis, autoimmune disease of the testis and ovary including autoimmune orchitis and oophoritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis such as autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis), or subacute thyroiditis, idiopathic hypothyroidism, Grave's disease, polyglandular syndromes such as autoimmune polyglandular syndromes (or polyglandular endocrinopathy syndromes), paraneoplastic syndromes, including neurologic paraneoplastic syndromes such as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, stiff-man or stiff-person syndrome, encephalomyelitis such as allergic encephalomyelitis or encephalomyelitis allergica and experimental allergic encephalomyelitis (EAE), myasthenia gravis such as thymoma-associated myasthenia gravis, cerebellar degeneration, neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome (OMS), and sensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome, autoimmune hepatitis, lupoid hepatitis, giant-cell hepatitis, autoimmune chronic active hepatitis, lymphoid interstitial pneumonitis (LIP), bronchiolitis obliterans (non-transplant) vs NSIP, Guillain-Barre syndrome, Berger's disease (IgA nephropathy), idiopathic IgA nephropathy, linear IgA dermatosis, acute febrile neutrophilic dermatosis, subcorneal pustular dermatosis, transient acantholytic dermatosis, cirrhosis such as primary biliary cirrhosis and pneumonocirrhosis, autoimmune enteropathy syndrome, Celiac or Coeliac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, amylotrophic lateral sclerosis (ALS; Lou Gehrig's disease), coronary artery disease, autoimmune ear disease such as autoimmune inner ear disease (AIED), autoimmune hearing loss, polychondritis such as refractory or relapsed or relapsing polychondritis, pulmonary alveolar proteinosis, Cogan's syndrome/nonsyphilitic interstitial keratitis, Bell's palsy, Sweet's disease/syndrome, rosacea autoimmune, zoster-associated pain, amyloidosis, a non-cancerous lymphocytosis, a primary lymphocytosis, which includes monoclonal B cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndrome, channelopathies including channelopathies of the CNS, autism, inflammatory myopathy, focal or segmental or focal segmental glomerulosclerosis (FSGS), endocrine opthalmopathy, uveoretinitis, chorioretinitis, autoimmune hepatological disorder, fibromyalgia, multiple endocrine failure, Schmidt's syndrome, adrenalitis, gastric atrophy, presenile dementia, demyelinating diseases such as autoimmune demyelinating diseases and chronic inflammatory demyelinating polyneuropathy, Dressler's syndrome, alopecia areata, alopecia totalis, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility, e.g., due to anti-spermatozoan antibodies, mixed connective tissue disease, Chagas' disease, rheumatic fever, recurrent abortion, farmer's lung, erythema multiforme, post-cardiotomy syndrome, Cushing's syndrome, bird-fancier's lung, allergic granulomatous angiitis, benign lymphocytic angitis, Alport's syndrome, alveolitis such as allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reaction, Sampter's syndrome, Caplan's syndrome, endocarditis, endomyocardial fibrosis, diffuse interstitial pulmonary fibrosis, interstitial lung fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilic faciitis, Shulman's syndrome, Felty's syndrome, cyclitis such as chronic cyclitis, heterochronic cyclitis, iridocyclitis (acute or chronic), or Fuch's cyclitis, Henoch-Schonlein purpura, SCID, sepsis, endotoxemia, post-vaccination syndromes, Evan's syndrome, autoimmune gonadal failure, Sydenham's chorea, post-streptococcal nephritis, thromboangitis ubiterans, thyrotoxicosis, tabes dorsalis, chorioiditis, giant-cell polymyalgia, chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, idiopathic nephritic syndrome, minimal change nephropathy, benign familial and ischemia-reperfusion injury, transplant organ reperfusion, retinal autoimmunity, aphthae, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Boeck's disease, enteritis allergica, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, febris rheumatica, Hamman-Rich's disease, sensoneural hearing loss, ileitis regionalis, leucopenia, transverse myelitis, primary idiopathic myxedema, ophthalmia symphatica, polyradiculitis acuta, pyoderma gangrenosum, acquired spenic atrophy, vitiligo, toxic-shock syndrome, conditions involving infiltration of T cells, leukocyte-adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes, diseases involving leukocyte diapedesis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathies, oophoritis, primary myxedema, autoimmune atrophic gastritis, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine failure, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome, primary sclerosing cholangitis, acute or chronic sinusitis, ethmoid, frontal, maxillary, or sphenoid sinusitis, an eosinophil-related disorder such as eosinophilia, pulmonary infiltration eosinophilia, eosinophilia-myalgia syndrome, Loffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, granulomas containing eosinophils, seronegative spondyloarthritides, polyendocrine autoimmune disease, sclerosing cholangitis, sclera, episclera, Bruton's syndrome, transient hypogammaglobulinemia of infancy, Wiskott-Aldrich syndrome, ataxia telangiectasia syndrome, angiectasis, autoimmune disorders associated with collagen disease, rheumatism, allergic hypersensitivity disorders, glomerulonephritides, reperfusion injury, ischemic re-perfusion disorder, lymphomatous tracheobronchitis, inflammatory dermatoses, dermatoses with acute inflammatory components, and autoimmune uveoretinitis (AUR). In some embodiments, the autoimmune disease or condition or T cell mediated inflammation can be neurodegeneration, e.g, Alzheimer's or Parksinson disease.

In some embodiments of any of the aspects, the autoimmune disease or condition, or T cell mediated inflammation can be type 1 diabetes, rheumatoid arthritis, multiple sclerosis, pemphigus, alopecia, lupus, vitiligo, or chronic fatigue syndrome.

As used herein, “malignant T cell condition” refers to a condition in which T cells display one or more of uncontrolled growth (i.e., division beyond normal limits), invasion (i.e., intrusion on and destruction of adjacent tissues), and metastasis (i.e., spread to other locations in the body via lymph or blood). Non-limiting examples of malignant T cell conditions include T cell cancers, lymphoma, leukemia, T cell acute lymphoblastic leukemia, and T cell lymphoblastic lymphoma.

The compositions and methods described herein can be administered to a subject to treat or prevent an autoimmune diseases or conditions; T cell mediated inflammation or immune response; or transplant rejection. In some embodiments, the methods described herein comprise administering an effective amount of compositions, CALs, CARs, or cells described herein to a subject in order to alleviate a symptom of an autoimmune diseases or conditions; T cell mediated inflammation or immune response; or transplant rejection. As used herein, “alleviating a symptom” is ameliorating any condition or symptom associated with the disease or condition. As compared with an equivalent untreated control, such reduction is by at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99% or more as measured by any standard technique. A variety of means for administering the compositions described herein to subjects are known to those of skill in the art. Such methods can include, but are not limited to oral, parenteral, intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, cutaneous, topical, injection, or intratumoral administration. Administration can be local or systemic.

The administration of the compositions contemplated herein may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. In a preferred embodiment, compositions are administered parenterally. The phrases “parenteral administration” and “administered parenterally” as used herein refers to modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravascular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intratumoral, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. In one embodiment, the compositions contemplated herein are administered to a subject by direct injection into a tumor, lymph node, or site of infection.

It can generally be stated that a pharmaceutical composition comprising the cells, e.g., T cells or CAL cells or CAR cells, described herein may be administered at a dosage of 10² to 10¹⁰ cells/kg body weight, preferably 10⁵ to 10⁶ cells/kg body weight, including all integer values within those ranges. The number of cells will depend upon the ultimate use for which the composition is intended as will the type of cells included therein. For uses provided herein, the cells are generally in a volume of a liter or less, can be 500 mLs or less, even 250 mLs or 100 mLs or less. Hence the density of the desired cells is typically greater than 10⁶ cells/ml and generally is greater than 10⁷ cells/ml, generally 10⁸ cells/ml or greater. The clinically relevant number of immune cells can be apportioned into multiple infusions that cumulatively equal or exceed 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10 ¹⁰, 10¹¹ or 10¹² cells. In some aspects of the present invention, particularly since all the infused cells will be redirected to a particular or specific target antigen, lower numbers of cells, in the range of 10⁶/kilogram (10⁶-10¹¹ per patient) may be administered. CAL and/or CAR expressing cell compositions may be administered multiple times at dosages within these ranges. The cells may be allogeneic, syngeneic, xenogeneic, or autologous to the patient undergoing therapy. If desired, the treatment may also include administration of mitogens (e.g., PHA) or lymphokines, cytokines, and/or chemokines (e.g., IFN-γ, IL-2, IL-12, TNF-alpha, IL-18, and TNF-beta, GM-CSF, IL-4, IL-13, Flt3-L, RANTES, MIP1α, etc.) as described herein to enhance induction of the immune response.

In some embodiments, the dosage can be from about 1×10⁵ cells to about 1×10⁸ cells per kg of body weight. In some embodiments, the dosage can be from about 1×10⁶ cells to about 1×10⁷ cells per kg of body weight. In some embodiments, the dosage can be about 1×10⁶ cells per kg of body weight. In some embodiments, one dose of cells can be administered. In some embodiments, the dose of cells can be repeated, e.g., once, twice, or more. In some embodiments, the dose of cells can be administered on, e.g., a daily, weekly, or monthly basis.

The dosage ranges for the agent, e.g., a CAL, CAR, cell, or composition described herein depend upon the potency, and encompass amounts large enough to produce the desired effect e.g., prevention of transplant rejection, reduction in inflammation, etc. The dosage should not be so large as to cause unacceptable adverse side effects. Generally, the dosage will vary with the age, condition, and sex of the patient and can be determined by one of skill in the art. The dosage can also be adjusted by the individual physician in the event of any complication. In some embodiments, the dosage ranges from 0.001 mg/kg body weight to 0.5 mg/kg body weight. In some embodiments, the dose range is from 5 μg/kg body weight to 100 μg/kg body weight. Alternatively, the dose range can be titrated to maintain serum levels between 1 μg/mL and 1000 μg/mL. For systemic administration, subjects can be administered a therapeutic amount, such as, e.g., 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, or more.

Administration of the doses recited above can be repeated. In some embodiments, the doses are given once a day, or multiple times a day, for example but not limited to three times a day. In some embodiments, the doses recited above are administered daily for several weeks or months. The duration of treatment depends upon the subject's clinical progress and responsiveness to therapy.

In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg. In some embodiments, the dose can be about 2 mg/kg. In some embodiments, the dose can be about 4 mg/kg. In some embodiments, the dose can be about 5 mg/kg. In some embodiments, the dose can be about 6 mg/kg. In some embodiments, the dose can be about 8 mg/kg. In some embodiments, the dose can be about 10 mg/kg. In some embodiments, the dose can be about 15 mg/kg. In some embodiments, the dose can be from about 100 mg/m² to about 700 mg/m². In some embodiments, the dose can be about 250 mg/m². In some embodiments, the dose can be about 375 mg/m². In some embodiments, the dose can be about 400 mg/m². In some embodiments, the dose can be about 500 mg/m².

In some embodiments, the dose can be administered intravenously. In some embodiments, the intravenous administration can be an infusion occurring over a period of from about 10 minutes to about 3 hours. In some embodiments, the intravenous administration can be an infusion occurring over a period of from about 30 minutes to about 90 minutes.

In some embodiments the dose can be administered about weekly. In some embodiments, the dose can be administered weekly. In some embodiments, the dose can be administered weekly for from about 12 weeks to about 18 weeks. In some embodiments the dose can be administered about every 2 weeks. In some embodiments the dose can be administered about every 3 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered about every 2 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered about every 3 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered intravenously about every 2 weeks. In some embodiments, the dose can be from about 2 mg/kg to about 15 mg/kg administered intravenously about every 3 weeks. In some embodiments, the dose can be from about 200 mg/m2 to about 400 mg/m2 administered intravenously about every week. In some embodiments, the dose can be from about 200 mg/m2 to about 400 mg/m2 administered intravenously about every 2 weeks. In some embodiments, the dose can be from about 200 mg/m2 to about 400 mg/m2 administered intravenously about every 3 weeks. In some embodiments, a total of from about 2 to about 10 doses are administered. In some embodiments, a total of 4 doses are administered. In some embodiments, a total of 5 doses are administered. In some embodiments, a total of 6 doses are administered. In some embodiments, a total of 7 doses are administered. In some embodiments, a total of 8 doses are administered. In some embodiments, the administration occurs for a total of from about 4 weeks to about 12 weeks. In some embodiments, the administration occurs for a total of about 6 weeks. In some embodiments, the administration occurs for a total of about 8 weeks. In some embodiments, the administration occurs for a total of about 12 weeks. In some embodiments, the initial dose can be from about 1.5 to about 2.5 fold greater than subsequent doses.

In some embodiments, the dose can be from about 1 mg to about 2000 mg. In some embodiments, the dose can be about 3 mg. In some embodiments, the dose can be about 10 mg. In some embodiments, the dose can be about 30 mg. In some embodiments, the dose can be about 1000 mg. In some embodiments, the dose can be about 2000 mg. In some embodiments, the dose can be about 3 mg given by intravenous infusion daily. In some embodiments, the dose can be about 10 mg given by intravenous infusion daily. In some embodiments, the dose can be about 30 mg given by intravenous infusion three times per week.

A therapeutically effective amount is an amount of an agent that is sufficient to produce a statistically significant, measurable change in, or prevent the occurrence of an autoimmune disease or condition; T cell mediated inflammation or immune response; transplant rejection; or GvHD. Such effective amounts can be gauged in clinical trials as well as animal studies.

An agent can be administered intravenously by injection or by gradual infusion over time. Given an appropriate formulation for a given route, for example, agents useful in the methods and compositions described herein can be administered intravenously, intranasally, by inhalation, intraperitoneally, intramuscularly, subcutaneously, intracavity, and can be delivered by peristaltic means, if desired, or by other means known by those skilled in the art. It is preferred that the compounds used herein are administered orally, intravenously or intramuscularly. Local administration, e.g., directly to the site of an organ or tissue transplant is also specifically contemplated.

Therapeutic compositions containing at least one agent can be conventionally administered in a unit dose, for example. The term “unit dose” when used in reference to a therapeutic composition refers to physically discrete units suitable as unitary dosage for the subject, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required physiologically acceptable diluent, i.e., carrier, or vehicle.

The compositions are administered in a manner compatible with the dosage formulation, and in a therapeutically effective amount. The quantity to be administered and timing depends on the subject to be treated, capacity of the subject's system to utilize the active ingredient, and degree of therapeutic effect desired.

In embodiments where the subject is administered a partial multi-component CAL and/or CAR cell and a recognition polypeptide, the partial multi-component CAL and/or CAR cell and a recognition polypeptide can be administered together or separately. In embodiments where the subject is separately administered a partial multi-component CAL and/or CAR cell and a recognition polypeptide each of the compositions can be administered, separately, according to any of the dosages and administration routes/routines described herein.

Precise amounts of active ingredient required to be administered depend on the judgment of the practitioner and are particular to each individual. However, suitable dosage ranges for systemic application are disclosed herein and depend on the route of administration. Suitable regimes for administration are also variable but are typified by an initial administration followed by repeated doses at one or more hour intervals by a subsequent injection or other administration. Alternatively, continuous intravenous infusion sufficient to maintain concentrations in the blood in the ranges specified for in vivo therapies are contemplated.

In some embodiments, the methods further comprise administering a composition, CAL, or CAR, or cell described herein along with one or more additional autoimmune, GvHD, or transplant rejection agents, biologics, drugs, or treatments as part of a combinatorial therapy. Exemplary treatments for transplant rejection or GvHD include but are not limited to, Immunosuppressive drugs, e.g., Cyclosporine (Neoral, Sandimmune, Gengraf, and Restasis), Tacrolimus (Prograf, Protopic, Astagraf XL, and Envarsus XR), Methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), Sirolimus (Rapamune), Mycophenolic acid (Myfortic and CellCept), Rituximab (Rituxan), etanercept (Enbrel), pentostatin (Nipent), ruxolitinib (Jakafi); Chemotherapies, e.g., Methotrexate (Trexall, Rasuvo, Rheumatrex, and Otrexup (PF)), antithymocyte globulin (Atgam, Thymoglobulin); Steroids, e.g., Prednisone (Deltasone, Rayos, and Prednisone Intensol), Methylprednisolone (Medrol, Solu-Medrol, and Depo-Medrol), budesonide (Entocort EC, Uceris); Antifungal, e.g., Posaconazole (Noxafil); Antiviral drugs, e.g., Acyclovir (Zovirax and Sitavig), Valacyclovir (Valtrex); and Antibiotics, e.g., Sulfamethoxazole/Trimethoprim (Bactrim, Sulfatrim, and Bactrim DS); Protease inhibitors, e.g. alphal-proteinase inhibitor (Zemaira); extracorporeal photopheresis; monoclonal antibodies (daclizumab (Zinbryta), basiliximab (Simulect)), Brentuximab vedotin (Adcetris), Alemtuzumab (Campath, Lemtrada), Tocilizumab (Actemra); infusion of mesenchymal stromal cells.

Exemplary treatments for autoimmune disease include but are not limited to, Insulin, e.g., Insulin glulisine (Apidra and Apidra SoloStar), Insulin detemir (Levemir and Levemir FlexTouch), Insulin aspart (NovoLog, Novolog Flexpen, and Novolog PenFill), Insulin lispro (Humalog and Humalog KwikPen), Insulin, Insulin glargine (Lantus, Lantus Solostar, and Toujeo SoloStar); Dietary supplement, e.g., glucose tablets; and Hormones, e.g., Glucagon (GlucaGen and Glucagon Emergency Kit (human)), antidiabetic agents (Metformin (D-Care DM2, Fortamet, Glucophage, Glucophage XR, Glumetza, Riomet), glucagon-like peptide-1 (GLP-1) receptor agonist (liraglutide (Saxenda; Victoza) or semaglutide (Ozempic) or sodium-glucose co-transporter 2 (SGLT2) inhibitor: empagliflozin (Jardiance), canagliflozin (Invokana); sulfonylureas: glipizide (GlipiZIDE XL, Glucotrol, Glucotrol XL); Meglitinide Analogs: repaglinide (Prandin); Thiazolidinedione: pioglitazone (Actos); dipeptidyl peptidase-4 (DPP-4) inhibitors: Sitagliptin (Januvia), Saxagliptin (Onglyza), Linagliptin (Tradjenta), Alogliptin (Nesina)

The efficacy of a given treatment, e.g., for an autoimmune diseases or conditions; T cell mediated inflammation or immune response; transplant rejection; or GvHD, can be determined by the skilled clinician. However, a treatment is considered “effective treatment,” as the term is used herein, if any one or all of the signs or symptoms of are altered in a beneficial manner or other clinically accepted symptoms are improved, or even ameliorated, e.g., by at least 10% following treatment with an agent as described herein. Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or described herein.

An effective amount for the treatment of a disease means that amount which, when administered to a mammal in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease. Efficacy of an agent can be determined by assessing physical indicators of, for example autoimmune disease (e.g., result of an ANA), T cell mediated inflammation or immune response, malignant T cell condition, transplant rejection (e.g., high fever, tenderness at transplant site, etc.), or GvHD (e.g., redness, pain, or other symptoms at transplant site). Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dosage can vary depending upon the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD50/ED50. Compositions and methods that exhibit large therapeutic indices are preferred. A therapeutically effective dose can be estimated initially from cell culture assays. Also, a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the active ingredient(s), which achieves a half-maximal inhibition of symptoms) as determined in cell culture, or in an appropriate animal model. Levels in plasma can be measured, for example, by immunoassay, various DNA detection technologies, or high performance liquid chromatography. The effects of any particular dosage can be monitored by a suitable bioassay, e.g., assay to assess reaction following transplant, level of inflammation, ANA measurement, among others. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization, or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or are described herein. Treatment includes any treatment of a disease in an individual or an animal (some non-limiting examples include a human or an animal) and includes: (1) inhibiting the disease, e.g., preventing a worsening of symptoms (e.g. pain or inflammation); or (2) relieving the severity of the disease, e.g., causing regression of symptoms. An effective amount for the treatment of a disease means that amount which, when administered to a subject in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease. Efficacy of an agent can be determined by assessing physical indicators of a condition or desired response, (e.g. a reduction of inflammation, etc.). It is well within the ability of one skilled in the art to monitor efficacy of administration and/or treatment by measuring any one of such parameters, or any combination of parameters. Efficacy can be assessed in animal models of a condition described herein, for example treatment of autoimmune disease, transplant rejection or GVHD. When using an experimental animal model, efficacy of treatment is evidenced when a statistically significant change in a marker is observed, e.g. inflammation.

In some embodiments, the technology described herein relates to a pharmaceutical composition comprising a CAL and/or CAR, or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR, or a multi-component CAL and/or CAR) as described herein, and optionally a pharmaceutically acceptable carrier. In some embodiments, the active ingredients of the pharmaceutical composition comprise a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR or a multi-component CAL and/or CAR) as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist essentially of a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a CAL and/or CAR or a multi-component CAL and/or CAR) as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist of a CAL and/or CAR or a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as described herein.

Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer solutions, solvents and/or dispersion media. The use of such carriers and diluents is well known in the art. Some non-limiting examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) serum component, such as serum albumin, HDL and LDL; (22) C2-C12 alcohols, such as ethanol; and (23) other non-toxic compatible substances employed in pharmaceutical formulations. Wetting agents, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservative and antioxidants can also be present in the formulation. The terms such as “excipient”, “carrier”, “pharmaceutically acceptable carrier” or the like are used interchangeably herein. In some embodiments, the carrier inhibits the degradation of the active agent as described herein.

In some embodiments, the pharmaceutical composition comprising a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as described herein can be a parenteral dose form. Since administration of parenteral dosage forms typically bypasses the patient's natural defenses against contaminants, parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. In addition, controlled-release parenteral dosage forms can be prepared for administration of a patient, including, but not limited to, DUROS®-type dosage forms and dose-dumping.

Suitable vehicles that can be used to provide parenteral dosage forms of a multi-component CAL and/or CAR (or portion thereof, or cell comprising a multi-component CAL and/or CAR) as disclosed within are well known to those skilled in the art. Examples include, without limitation: sterile water; water for injection USP; saline solution; glucose solution; aqueous vehicles such as but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate. Compounds that alter or modify the solubility of a pharmaceutically acceptable salt of an active ingredient can also be incorporated into the parenteral dosage forms of the disclosure, including conventional and controlled-release parenteral dosage forms.

Pharmaceutical compositions can also be formulated to be suitable for oral administration, for example as discrete dosage forms, such as, but not limited to, tablets (including without limitation scored or coated tablets), pills, caplets, capsules, chewable tablets, powder packets, cachets, troches, wafers, aerosol sprays, or liquids, such as but not limited to, syrups, elixirs, solutions or suspensions in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion, or a water-in-oil emulsion. Such compositions contain a predetermined amount of the pharmaceutically acceptable salt of the disclosed compounds, and may be prepared by methods of pharmacy well known to those skilled in the art. See generally, Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams, and Wilkins, Philadelphia Pa. (2005).

Conventional dosage forms generally provide rapid or immediate drug release from the formulation. Depending on the pharmacology and pharmacokinetics of the drug, use of conventional dosage forms can lead to wide fluctuations in the concentrations of the drug in a patient's blood and other tissues. These fluctuations can impact a number of parameters, such as dose frequency, onset of action, duration of efficacy, maintenance of therapeutic blood levels, toxicity, side effects, and the like. Advantageously, controlled-release formulations can be used to control a drug's onset of action, duration of action, plasma levels within the therapeutic window, and peak blood levels. In particular, controlled- or extended-release dosage forms or formulations can be used to ensure that the maximum effectiveness of a drug is achieved while minimizing potential adverse effects and safety concerns, which can occur both from under-dosing a drug (i.e., going below the minimum therapeutic levels) as well as exceeding the toxicity level for the drug. In some embodiments, the composition can be administered in a sustained release formulation.

Controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled release counterparts. Ideally, the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled-release formulations include: 1) extended activity of the drug; 2) reduced dosage frequency; 3) increased patient compliance; 4) usage of less total drug; 5) reduction in local or systemic side effects; 6) minimization of drug accumulation; 7) reduction in blood level fluctuations; 8) improvement in efficacy of treatment; 9) reduction of potentiation or loss of drug activity; and 10) improvement in speed of control of diseases or conditions. Kim, Cherng-ju, Controlled Release Dosage Form Design, 2 (Technomic Publishing, Lancaster, Pa.: 2000).

Most controlled-release formulations are designed to initially release an amount of drug (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release other amounts of drug to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, ionic strength, osmotic pressure, temperature, enzymes, water, and other physiological conditions or compounds.

A variety of known controlled- or extended-release dosage forms, formulations, and devices can be adapted for use with the salts and compositions of the disclosure. Examples include, but are not limited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,733,566; and 6,365,185 B1; each of which is incorporated herein by reference. These dosage forms can be used to provide slow or controlled-release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems (such as OROS® (Alza Corporation, Mountain View, Calif. USA)), or a combination thereof to provide the desired release profile in varying proportions.

Described herein is a therapy called “CAL-cell therapy”, which seeks to help immune killer cells recognize autoreactive and alloreactive T cells. This is accomplished by genetically altering an immune cell so that it expresses a chimeric antigen ligand (CAL). The CAL is an altered ligand, in which the natural recognition portion is removed and replaced with a synthetic recognition portion, (including all synthetic or natural peptide MHC complexes) that is designed to more effectively recognize the autoreactive and alloreactive T cells by very specifically detecting the presence of a T cell receptor unique to the autoreactive and alloreactive T cells. These CAL immune cells are then given to a patient. Inside the patient, their synthetic CAL ligand molecules will bind to the autoreactive and alloreactive T cells and in the act of that binding, activate the killer immune cells, resulting in the engineered CAL-immune cells attacking the pathologic autoreactive and alloreactive T-cells to eliminate or reduce allo- or autoreactive immune response. This has particular applications for autoimmune conditions, cell-, tissue-, organ-transplants, and Graft vs. Host Disease (GvHD).

This invention is based, in part, on the finding that an engineered polypeptide presented herein can recognize and bind to the specific T cell receptor on the disease-causing T cells, deleting said T cells, e.g., eliminating or reducing autoimmune diseases or conditions; T cell mediated inflammation or immune response; and cell, tissue, organ transplants and Graft vs. Host Disease (GvHD). In some embodiments, the engineered polypeptide is composed of a peptide-major histocompatibility complex (pMHC) (e.g. as a monomer, oligomer, or multimer) as the recognition site for the TCR of an allogeneic or an autoreactive T cell. In some embodiments, the pMHC is one of the complexes described herein, e.g., see Tables 5 and 6.

In some embodiments, the engineered polypeptide is composed of a pMHC conjugated to a FITC, PE, or other biomolecular interaction domain described herein. The engineered polypeptide can also be considered as an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to a biomolecular interaction domain. The CAL technology described herein can target T cell clones in an antigen specific manner and is not dependent on any specific CAR construct to exhibit killing effect.

In some embodiments, the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as the recognition site fused to signaling domains from T cell receptors. Further provided herein is a split version of this CAR system, in which the CAR is composed of two pieces. The first piece is a universal CAR (also referred to herein as a Uni CAL) with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain, e.g., that is specific to each disease state. The second piece is an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to the cognate biomolecular interaction domain.

In some embodiments, the engineered polypeptide is a CAR composed of a peptide-HLA (e.g., as a monomer, oligomer, or multimer) as the recognition site fused to signaling domains from T cell receptors. Further provided herein is a split version of this CAR system, in which the CAR is composed of two pieces. The first piece is a universal CAR (also referred to herein as a Uni CAL) with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain, e.g., that is specific to each disease state. The second piece is an adaptor molecule (referred to herein as Chimeric Antigen Ligand (CAL)) composed of a TCR recognition domain (e.g., peptide-HLA monomers, oligomers, or multimers) fused to the cognate biomolecular interaction domain. It is noted that protein interaction domains are a type of biomolecular interaction domains and where one is specified herein, the other may always be substituted. The CAL technology described herein can target T cells clones in an antigen specific manner and is not dependent on any specific CAR construct to exhibit killing effect.

Accordingly, one aspect presented herein provides a composition comprising (a) chimeric antigen ligand or a TCR recognition domain; and one or both of (a) an intracellular signaling domain; and (b) a first-type protein interaction domain.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a chimeric antigen ligand or TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; wherein the first-type and second-type protein interaction domains bind specifically to each other.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the first-type and third-type protein interaction domains bind specifically to each other.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; and (b) a signaling polypeptide comprising a second-type protein interaction domain and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a third-type protein interaction domain; wherein the second-type and third-type protein interaction domains compete for binding to the first-type protein interaction domain.

In one embodiment of any aspect, the third-type protein interaction domain and first-type protein interaction domain have a higher affinity for each other than the second-type protein interaction domain and first-type protein interaction domain.

Another aspect provided herein provides a composition comprising (a) a first polypeptide comprising a TCR recognition domain and a first-type protein interaction domain; (b) a signaling polypeptide comprising a second-type protein interaction domain, a fourth-type protein interaction domain, and an intracellular signaling domain; and (c) a recognition polypeptide comprising a second recognition domain and a fifth-type protein interaction domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

In one embodiment of any aspect, the fourth-type protein interaction domain and fifth-type protein interaction domain have a weaker affinity than the second-type protein interaction domain and first-type protein interaction domain.

In one embodiment of any aspect, the first polypeptide further comprises a sixth-type protein interaction domain and the recognition polypeptide further comprises a seventh-type protein interaction domain which bind specifically to each other.

In one embodiment of any aspect, the second recognition domain is specific for a target that is not recognized by the TCR recognition domain.

In one embodiment of any aspect, the second recognition domain is specific for a target that is found on a healthy and/or non-target cell and not on a diseased and/or target cell.

TCR recognition domains may not comprise a single polypeptide, but rather comprise two or more polypeptides and may additionally even comprise non-polypeptides. For example, a single MHC class II tetramer TCR recognition domain can comprise 4 biotin small molecules and 16 polypeptides (4 peptides, 4 MHC class II alpha chains, 4 MHC class II beta chains, and 4 streptavidin proteins. Other types of TCR recognition domains may additionally comprise other non-polypeptide molecules (e.g., MHC dextramers contain a polysaccharide backbone to which the MI-ICs are anchored to). In some embodiments, a composition described herein can comprise multiple copies or instances of a TCR recognition domain(s), e.g. the TCR recognition domain can be a mulitmer, or oligomer. In some embodiments, a composition described herein can comprise multiple copies or instances of a first polypeptide as described herein. In some embodiments, the first polypeptide comprises the entire TCR recognition domain. In some embodiments, the TCR recognition domain comprises at least two separate polypeptide sequences, the first polypeptide comprises at least one of the separate polypeptide sequences of the TCR recognition domain, and the first polypeptide is bound to or complexed with a second or further polypeptide sequences of the TCR recognition domain to form a TCR recognition domain.

In one embodiment of any aspect, the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; or a MHC-peptide fusion.

In one embodiment of any aspect, the peptide is a human, non-human, or synthetic/engineered peptide. The peptides can further comprise non-proteinaceous motifs, modifications, or domains, e.g., they can comprise glycosylation and/or lipids. In one embodiment of any aspect, the peptide is a Minor Histocompatibility Antigen (MiHA).

In one embodiment of any aspect, the MHC is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.

In one embodiment of any aspect, the protein interaction domains are found on an extracellular portion of the respective polypeptides.

In one embodiment of any aspect, (a) the protein interaction domain(s) is a leucine zipper, or any binding pair of protein interaction domains are collectively a pair of leucine zippers; (b) the protein interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of protein interaction domains are collectively a BZip (RR) and a AZip (EE); (c) the protein interaction domain(s) is a PSD95-D1g1-zo-1 (PDZ) domain; (d) the protein interaction domain(s) is a streptavidin and/or a streptavidin binding protein (SBP) or any binding pair of protein interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP); (e) the protein interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding protein (FKBP); (f) the protein interaction domain(s) is a cyclophilin-Fas fusion protein (CyP-Fas) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a cyclophilin-Fas fusion protein (CyP-Fas) and a FK506 binding protein (FKBP); (g) the protein interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding protein (FKBP) or any binding pair of protein interaction domains are collectively a calcineurin A (CNA) and a FK506 binding protein (FKBP); (h) the protein interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of protein interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); (i) the protein interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of protein interaction domains are collectively a Snap-tag and a Halo tag; (j) the protein interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of protein interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); (k) the protein interaction domain(s) is a PYL and/or a ABI, or any binding pair of protein interaction domains are collectively a PYL and a ABI; (l) the protein interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of protein interaction domains are collectively a nucleotide tag and a zinc finger domain; (m) wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags; (n) the protein interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding protein or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or (o) the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein.

In one embodiment of any aspect, the nucleotide tag is a DNA tag or dsDNA tag.

In one embodiment of any aspect, the intracellular signaling domain is a signaling domain from a protein selected from the group consisting of: TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; and ZAP70.

In one embodiments of any aspect, a cell comprising and/or expressing a comprising a composition comprising a TCR recognition domain and an intracellular signaling domain further comprises a TCR signaling-responsive promoter operatively linked to a payload transgene. Such embodiments permit transgene payload expression specifially to and/or in the vicinity of a targeted T cell. Suitable promoters and transgene are known in the art, e.g., those promoters and transgenes used in “TRUCK CAR” technology. An exemplary promoter is a NFAT-sensitive promoter. Exemplary transgene payloads can include checkpoint inhibitors (e.g., CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]) or proinflammatory cytokines (e.g., IL-2, IL-12, etc). In our case this will be used for in-situ targeting of a patient's anticancer T-cells to very specifically and locally deliver activating agents like one or more checkpoint inhibitors (CTLA-4, [Ipilimumab, Tremelimumab] or PD-1 [Nivolumab, Pembrolizumab, Pidilizumab]) and/or proinflammatory cytokines, (e.g. IL-2, IL-12, etc.) that can push them to expansion and effector phenotype. With this targeted delivery, it is expected to see more efficacy without the systemic side effects caused by checkpoint inhibitors and cytokines administered systemically. Further, combination therapy with multiple different agents is possible with the described technolog as there is minimal systemic side effect due to paracrine delivery. Further description of suitable promoters, payloads, as well as how to make and use TRUCK technology is described, e.g., Peterson et al. Front. Oncol. 2019 9:69; Chmielewski et al. Advances in Cell and Gene Therapy 2020 3:e84; Chimielewski et al. Expert opinion Biol Ther 2015 15:1145-54; and Chimieleski et al. Immunol Rev 2014 257:83-90; each of which is incorporated by reference herein in its entirety. In some embodiments, the cell can be allogeneic, e.g., and engineered once and given as pulse therapy. In some embodiments, the cell can be a T cell or any other cell type described herein, e.g., a NK cell. Exemplary, non-limiting proinflammatory cytokines include IFNs, IFN-γ, TNFα, TGF-β, IL-1β, IL-6, IL-4, IL-10, IL-13, IL-2, IL-12, IL-15, and IL-27.

A promoter can be said to drive expression or drive transcription of the nucleic acid sequence that it regulates. The phrases “operably linked,” “operatively positioned,” “operatively linked,” “under control,” and “under transcriptional control” indicate that a promoter is in a correct functional location and/or orientation in relation to a nucleic acid sequence it regulates to control transcriptional initiation and/or expression of that sequence.

In some embodiments, a composition comprising a TCR recognition domain and an biomolecular interaction domain is a soluble molecule and/or soluble complex.

Another aspect provided herein is a cell comprising and/or expressing the composition of any of the compositions described herein.

In one embodiment of any aspect, the TCR recognition domain comprises a MHC allogeneic to the cell comprising and/or expressing the composition. In one embodiment of any aspect, the TCR recognition domain comprises a MHC allogeneic to the cell that the TCR originated from.

In one embodiment of any aspect, the TCR recognition domain comprises a peptide allogeneic to the cell comprising and/or expressing the composition. In one embodiment of any aspect, the TCR recognition domain comprises a peptide allogeneic to the cell that the TCR originated from.

In one embodiment of any aspect, the cell is a dendritic cell (CAL DC), a T cell (e.g., effector, regulatory, etc.) (CAL-T); regulatory T cell, effector T cell, natural killer cell (CAL NK), or any other myeloid cell. In one embodiment of any aspect, the cell is engineered to express the polypeptide(s) of the composition. In one embodiment of any aspect, the cell is engineered to express the signaling polypeptide of the composition. In one embodiment of any aspect, the cell is further engineered to knockout the native MHCI/II. In one embodiment of any aspect, the cell is further engineered to lack cell surface expression of native MHCI/II.

Another aspect provided herein is a chimeric antigen receptor (CAR) comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) an intracellular signaling domain.

Another aspect provided herein is a composition comprising a first polypeptide comprising: (a) an anti-CD127 and/or anti-CD45RO recognition domain; (b) a first-type protein interaction domain; and a second polypeptide comprising (a) a second-type protein interaction domain; and (b) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.

Another aspect provided herein is a composition comprising a first polypeptide comprising (a) an anti-CD127 recognition domain; (b) a first-type protein interaction domain; a second polypeptide comprising (a) an anti-CD45RO recognition domain; (b) a fifth-type protein interaction domain; and a third polypeptide comprising (a) a second-type and a fourth-type protein interaction domain; and (b) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

Another aspect provided herein is a cell comprising any of the CARs described herein, or any of the compositions described herein.

Another aspect provided herein is a method of preventing and/or treating an autoimmune diseases or conditions or T cell mediated inflammation or immune response; or treating or preventing transplant rejection or GvHD in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein. Another aspect provided herein is a method of preventing and/or treating a malignant T cell condition in a subject in need thereof, the method comprising administering to the subject any of the compositions and/or cells described herein.

In various embodiments of any aspect, the TCR recognition domain comprises a MHC allogeneic to the subject, a MHC autologous to the transplant cells, a peptide allogeneic to the subject, or a peptide autologous to the transplant cells.

In one embodiment of any aspect, the transplant is vascularized composite allotransplantation (VCA).

In one embodiment of any aspect, the autoimmune disease is type 1 diabetes, multiple sclerosis, rheumatoid arthritis, or scleroderma.

One aspect of the embodiments provided herein is a CAR T cell that targets a CD127+/CD45RO+ T cell. In one embodiment, the CD127+/CD45RO+ T cell is a CD127+/CD45RO+ memory T cell. In one embodiment, the CD127+/CD45RO+ T cell is an alloreactive CD127+/CD45RO+ T cell.

Accordingly, one aspect herein provides a CAR comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) intracellular signaling domain. Further provided herein is a composition comprising (a) an anti-CD127 and/or anti-CD45RO recognition domain; and (b) intracellular signaling domain.

Another aspect of the provides a composition comprising a first polypeptide comprising: (a) an anti-CD127 and/or anti-CD45RO recognition domain; (b) a first-type protein interaction domain; and a second polypeptide comprising: (c) a second-type protein interaction domain; and (d) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other.

Another aspect of the provides a composition comprising a first polypeptide comprising: (a) an anti-CD127 recognition domain; (b) a first-type protein interaction domain; a second polypeptide comprising: (c) an anti-CD45RO recognition domain; (d) a fifth-type protein interaction domain; and a third polypeptide comprising: (e) a second-type and a fourth-type protein interaction domain; and (f) an intracellular signaling domain; wherein the first-type protein interaction domain and the second-type protein interaction domain bind specifically to each other; and wherein the fourth-type protein interaction domain and the fifth-type protein interaction domain bind specifically to each other.

In one embodiment, the anti-CD127 recognition domain recognizes and binds to the sequence of CD127 on a CD127+ cell, e.g., a CD127+/CD45RO+ T cell. As used herein, “CD127”, also referred to as Interleukin 7 receptor a (IL7Ra), ILRA, IL7RA, CDW127, or IL-7R-alpha, is a cell surface receptor that has been shown to have a role in V(D)J recombination during lymphocyte development. Defects in CD127 have been associated with severe combined immunodeficiency (SCID). Sequences for CD127 are known for a number of species, e.g., human CD127 (NCBI Gene ID: 3575), mRNA (NCBI Ref Seq: NM_002185.5), and polypeptide (NCBI Ref Seq: NP_002176.2). CD127 refers to all naturally occurring variants or isoforms of CD127. In one embodiment, the CD127 polypeptide sequence is presented in SEQ ID NO: 1. In some embodiments of any of the aspects, the CD127 polypeptide can be an ortholog, variant, and/or allele of SEQ ID NO: 1.

(SEQ ID NO: 1)   1 MTILGTTFGM VFSLLQVVSG ESGYAQNGDL EDAELDDYSF      SCYSQLEVNG SQHSLTCAFE  61 DPDVNITNLE FEICGALVEV KCLNFRKLQE IYFIETKKFL      LIGKSNICVK VGEKSLTCKK 121 IDLTTIVKPE APFDLSVVYR EGANDFVVTF NTSHLQKKYV      KVLMHDVAYR QEKDENKWTH 181 VNLSSTKLTL LQRKLQPAAM YEIKVRSIPD HYFKGFWSEW      SPSYYFRTPE INNSSGEMDP 241 ILLTISILSF FSVALLVILA CVLWKKRIKP IVWPSLPDHK      KTLEHLCKKP RKNLNVSFNP 301 ESFLDCQIHR VDDIQARDEV EGFLQDTFPQ QLEESEKQRL      GGDVQSPNCP SEDVVITPES 361 FGRDSSLTCL AGNVSACDAP ILSSSRSLDC RESGKNGPHV      YQDLLLSLGT TNSTLPPPFS 421 LQSGILTLNP VAQGQPILTS LGSNQEEAYV TMSSFYQNQ 

In one embodiment, the anti-CD45RO recognition domain recognizes and binds to the sequence of CD45RO on a CD45RO+ cell, e.g., a CD127+/CD45RO+ T cell. As used herein, “CD45RO”, also referred to as PTPRC, LCA, LYS, B220, CD45, L-CA, T200, CD45R, and GP180, is to a cell surface signaling molecule that been shown to be an essential regulator of T- and B-cell antigen receptor signaling. Sequences for CD45RO are known for a number of species, e.g., human CD45RO (NCBI Gene ID: 5788), mRNA (NCBI Ref Seq: NM_001267798.2), and polypeptide (NCBI Ref Seq: NP_001254727.1). CD45RO refers to all naturally occurring variants or isoforms of CD45RO. In one embodiment, the CD45RO polypeptide sequence is presented in SEQ ID NO: 2. In some embodiments of any of the aspects, the CD45RO polypeptide can be an ortholog, variant, and/or allele of SEQ ID NO: 2.

(SEQ ID NO: 2)  1 MTMYLWLKLL AFGFAFLDTE VFVTGQSPTP SPTGHLQAEE     QGSQSKSPNL KSREADSSAF 61 SWWPKAREPL TNHWSKSKSP KAEELGV 

For convenience, the meaning of some terms and phrases used in the specification, examples, and appended claims, are provided below. Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is an apparent discrepancy between the usage of a term in the art and its definition provided herein, the definition provided within the specification shall prevail.

For convenience, certain terms employed herein, in the specification, examples and appended claims are collected here.

The terms “decrease”, “reduced”, “reduction”, or “inhibit” are all used herein to mean a decrease by a statistically significant amount. In some embodiments, “reduce,” “reduction” or “decrease” or “inhibit” typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, “reduction” or “inhibition” does not encompass a complete inhibition or reduction as compared to a reference level. “Complete inhibition” is a 100% inhibition as compared to a reference level. A decrease can be preferably down to a level accepted as within the range of normal for an individual without a given disorder.

The terms “increased”, “increase”, “enhance”, or “activate” are all used herein to mean an increase by a statically significant amount. In some embodiments, the terms “increased”, “increase”, “enhance”, or “activate” can mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level. In the context of a marker or symptom, a “increase” is a statistically significant increase in such level.

As used herein, a “subject” means a human or animal. Usually the animal is a vertebrate such as a swine, primate, rodent, domestic animal or game animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits and hamsters. Domestic and game animals include cows, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon. In some embodiments, the subject is a mammal, e.g., a primate, e.g., a human. The terms, “individual,” “patient” and “subject” are used interchangeably herein.

Preferably, the subject is a mammal. The mammal can be a human, non-human swine, primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of autoimmune diseases or conditions; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. A subject can be male or female.

A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment (e.g. an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD) or one or more complications related to such a condition, and optionally, have already undergone treatment for an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or the one or more complications related to an autoimmune disease, transplant rejection, or GvHD. Alternatively, a subject can also be one who has not been previously diagnosed as having an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or one or more complications related to an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. For example, a subject can be one who exhibits one or more risk factors for an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or one or more complications related to an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD or a subject who does not exhibit risk factors.

A “subject in need” of treatment for a particular condition can be a subject having that condition, diagnosed as having that condition, or at risk of developing that condition, e.g., an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD.

In some embodiments, a nucleic acid encoding a CAL, CAR, a multi-component CAL and/or CAR or portion thereof as described herein is comprised by a vector. In some of the aspects described herein, a nucleic acid sequence encoding a multi-component CAL and/or CAR, or portion thereof as described herein, or any module thereof, is operably linked to a vector. The term “vector”, as used herein, refers to a nucleic acid construct designed for delivery to a host cell or for transfer between different host cells. As used herein, a vector can be viral or non-viral. The term “vector” encompasses any genetic element that is capable of replication when associated with the proper control elements and that can transfer gene sequences to cells. A vector can include, but is not limited to, a cloning vector, an expression vector, a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc.

As used herein, the term “expression vector” refers to a vector that directs expression of an RNA or polypeptide from sequences linked to transcriptional regulatory sequences on the vector. The sequences expressed will often, but not necessarily, be heterologous to the cell. An expression vector may comprise additional elements, for example, the expression vector may have two replication systems, thus allowing it to be maintained in two organisms, for example in human cells for expression and in a prokaryotic host for cloning and amplification. The term “expression” refers to the cellular processes involved in producing RNA and proteins and as appropriate, secreting proteins, including where applicable, but not limited to, for example, transcription, transcript processing, translation and protein folding, modification and processing. “Expression products” include RNA transcribed from a gene, and polypeptides obtained by translation of mRNA transcribed from a gene. The term “gene” means the nucleic acid sequence that is transcribed (DNA) to RNA in vitro or in vivo when operably linked to appropriate regulatory sequences. The gene may or may not include regions preceding and following the coding region, e.g. 5′ untranslated (5′UTR) or “leader” sequences and 3′ UTR or “trailer” sequences, as well as intervening sequences (introns) between individual coding segments (exons).

As used herein, the term “viral vector” refers to a nucleic acid vector construct that includes at least one element of viral origin and has the capacity to be packaged into a viral vector particle. The viral vector can contain the nucleic acid encoding a CAL and/or CAR described herein, e.g., a multi-component CAL and/or CAR, or portion thereof as described herein in place of non-essential viral genes. The vector and/or particle may be utilized for the purpose of transferring any nucleic acids into cells either in vitro or in vivo. Numerous forms of viral vectors are known in the art.

By “recombinant vector” is meant a vector that includes a heterologous nucleic acid sequence, or “transgene” that is capable of expression in vivo or in the transduced cells. It should be understood that the vectors described herein can, in some embodiments, be combined with other suitable compositions and therapies. In some embodiments, the vector is episomal. The use of a suitable episomal vector provides a means of maintaining the nucleotide of interest in the subject in high copy number extra chromosomal DNA thereby eliminating potential effects of chromosomal integration.

As used herein, the term “nucleic acid” or “nucleic acid sequence” refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analog thereof. The nucleic acid can be either single-stranded or double-stranded. A single-stranded nucleic acid can be one nucleic acid strand of a denatured double-stranded DNA. Alternatively, it can be a single-stranded nucleic acid not derived from any double-stranded DNA. In one aspect, the nucleic acid can be DNA. In another aspect, the nucleic acid can be RNA, e.g., single-stranded or double-stranded RNA. Suitable nucleic acid molecules are DNA, including genomic DNA or cDNA. Other suitable nucleic acid molecules are RNA, including mRNA.

As used herein, the terms “protein” and “polypeptide” are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues. The terms “protein”, and “polypeptide” refer to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogs, regardless of its size or function. “Protein” and “polypeptide” are often used in reference to relatively large polypeptides, whereas the term “peptide” is often used in reference to small polypeptides, but usage of these terms in the art overlaps. The terms “protein” and “polypeptide” are used interchangeably herein when referring to a gene product and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogs of the foregoing.

As used herein an “antibody” refers to IgG, IgM, IgA, IgD or IgE molecules or antigen-specific antibody fragments thereof (including, but not limited to, a Fab, F(ab′)2, Fv, disulphide linked Fv, scFv, single domain antibody, closed conformation multispecific antibody, disulphide-linked scfv, diabody), whether derived from any species that naturally produces an antibody, or created by recombinant DNA technology; whether isolated from serum, B-cells, hybridomas, transfectomas, yeast or bacteria.

As described herein, an “antigen” is a molecule that is bound by a binding site on an antibody agent. Typically, antigens are bound by antibody ligands and are capable of raising an antibody response in vivo. An antigen can be a polypeptide, protein, nucleic acid or other molecule or portion thereof. The term “antigenic determinant” refers to an epitope on the antigen recognized by an antigen-binding molecule, and more particularly, by the antigen-binding site of said molecule.

As used herein, the term “antibody reagent” refers to a polypeptide that includes at least one immunoglobulin variable domain or immunoglobulin variable domain sequence and which specifically binds a given antigen. An antibody reagent can comprise an antibody or a polypeptide comprising an antigen-binding domain of an antibody. In some embodiments, an antibody reagent can comprise a monoclonal antibody or a polypeptide comprising an antigen-binding domain of a monoclonal antibody. For example, an antibody can include a heavy (H) chain variable region (abbreviated herein as VH), and a light (L) chain variable region (abbreviated herein as VL). In another example, an antibody includes two heavy (H) chain variable regions and two light (L) chain variable regions. The term “antibody reagent” encompasses antigen-binding fragments of antibodies (e.g., single chain antibodies, Fab and sFab fragments, F(ab′)2, Fd fragments, Fv fragments, scFv, and domain antibodies (dAb) fragments (see, e.g. de Wildt et al., Eur J. Immunol. 1996; 26(3):629-39; which is incorporated by reference herein in its entirety)) as well as complete antibodies. An antibody can have the structural features of IgA, IgG, IgE, IgD, IgM (as well as subtypes and combinations thereof). Antibodies can be from any source, including mouse, rabbit, pig, rat, and primate (human and non-human primate) and primatized antibodies. Antibodies also include midibodies, humanized antibodies, chimeric antibodies, and the like.

The VH and VL regions can be further subdivided into regions of hypervariability, termed “complementarity determining regions” (“CDR”), interspersed with regions that are more conserved, termed “framework regions” (“FR”). The extent of the framework region and CDRs has been precisely defined (see, Kabat, E. A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, and Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917; which are incorporated by reference herein in their entireties). Each VH and VL is typically composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

The terms “antigen-binding fragment” or “antigen-binding domain”, which are used interchangeably herein are used to refer to one or more fragments of a full length antibody that retain the ability to specifically bind to a target of interest. Examples of binding fragments encompassed within the term “antigen-binding fragment” of a full length antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment including two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546; which is incorporated by reference herein in its entirety), which consists of a VH or VL domain; and (vi) an isolated complementarity determining region (CDR) that retains specific antigen-binding functionality.

As used herein, the term “specific binding” refers to a chemical interaction between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with greater specificity and affinity than it binds to a third entity which is a non-target. In some embodiments, specific binding can refer to an affinity of the first entity for the second target entity which is at least 10 times, at least 50 times, at least 100 times, at least 500 times, at least 1000 times or greater than the affinity for the third nontarget entity. A reagent specific for a given target is one that exhibits specific binding for that target under the conditions of the assay being utilized. In some embodiments, binding described herein can be preferential binding, e.g., binding between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with at least 2 times greater specificity and affinity than it binds to a third entity which is a non-target.

Additionally, and as described herein, a recombinant humanized antibody can be further optimized to decrease potential immunogenicity, while maintaining functional activity, for therapy in humans. In this regard, functional activity means a polypeptide capable of displaying one or more known functional activities associated with a recombinant antibody or antibody reagent thereof as described herein. Such functional activities include, e.g. the ability to bind to a target.

As used herein, the terms “treat,” “treatment,” “treating,” or “amelioration” refer to therapeutic treatments, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a condition associated with a disease or disorder, e.g. autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. The term “treating” includes reducing or alleviating at least one adverse effect or symptom of a condition, disease or disorder associated with a, e.g. autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of a disease is reduced or halted. That is, “treatment” includes not just the improvement of symptoms or markers, but also a cessation of, or at least slowing of, progress or worsening of symptoms compared to what would be expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, remission (whether partial or total), and/or decreased mortality, whether detectable or undetectable. The term “treatment” of a disease also includes providing relief from the symptoms or side-effects of the disease (including palliative treatment).

As used herein, the term “pharmaceutical composition” refers to the active agent in combination with a pharmaceutically acceptable carrier e.g. a carrier commonly used in the pharmaceutical industry. The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

As used herein, the term “administering,” refers to the placement of an agent, e.g., a CAL, CAR, composition, or cell as disclosed herein into a subject by a method or route which results in at least partial delivery of the agent at a desired site. Pharmaceutical compositions comprising the compounds disclosed herein can be administered by any appropriate route which results in an effective treatment in the subject.

Immune checkpoint inhibitors inhibit one or more immune checkpoint proteins. The immune system has multiple inhibitory pathways that are critical for maintaining self-tolerance and modulating immune responses. For example, in T-cells, the amplitude and quality of response is initiated through antigen recognition by the T-cell receptor and is regulated by immune checkpoint proteins that balance co-stimulatory and inhibitory signals. In some embodiments of any of the aspects, a subject or patient is treated with at least one inhibitor of an immune checkpoint protein. As used herein, “immune checkpoint protein” refers to a protein which, when active, exhibits an inhibitory effect on immune activity, e.g., T cell activity. Exemplary immune checkpoint proteins can include PD-1 (e.g., NCBI Gene ID: 5133); PD-L1 (e.g., NCBI Gene ID: 29126); PD-L2 (e.g., NCBI Gene ID: 80380); TIM-3 (e.g., NCBI Gene ID: 84868); CTLA4 (e.g., NCBI Gene ID: 1493); TIGIT (e.g., NCBI Gene ID: 201633); KIR (e.g., NCBI Gene ID: 3811); LAG3 (e.g., NCBI Gene ID: 3902); DD1-α (e.g., NCBI Gene ID: 64115); A2AR (e.g., NCBI Gene ID: 135); B7-H3 (e.g., NCBI Gene ID: 80381); B7-H4 (e.g., NCBI Gene ID: 79679); BTLA (e.g., NCBI Gene ID: 151888); IDO (e.g., NCBI Gene ID: 3620); TDO (e.g., NCBI Gene ID: 6999); HVEM (e.g., NCBI Gene ID: 8764); GALS (e.g., NCBI Gene ID: 3965); 2B4 (belongs to the CD2 family of molecules and is expressed on all NK, γδ, and memory CD8+ (αβ) T cells) (e.g., NCBI Gene ID: 51744); CD160 (also referred to as BY55) (e.g., NCBI Gene ID: 11126); and various B-7 family ligands. B7 family ligands include, but are not limited to, B7-1, B7-2, B7-DC, B7-H1, B7-H2, B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7.

Non-limiting examples of immune checkpoint inhibitors (with checkpoint targets and manufacturers noted in parantheses) can include: MGA271 (B7-H3: MacroGenics); ipilimumab (CTLA-4; Bristol Meyers Squibb); pembrolizumab (PD-1; Merck); nivolumab (PD-1; Bristol Meyers Squibb); atezolizumab (PD-L1; Genentech); galiximab (B7.1; Biogen); IMP321 (LAG3: Immuntep); BMS-986016 (LAG3; Bristol Meyers Squibb); SMB-663513 (CD137; Bristol-Meyers Squibb); PF-05082566 (CD137; Pfizer); IPH2101 (KIR; Innate Pharma); KW-0761 (CCR4; Kyowa Kirin); CDX-1127 (CD27; CellDex); MEDI-6769 (Ox40; MedImmune); CP-870,893 (CD40; Genentech); tremelimumab (CTLA-4; Medimmune); pidilizumab (PD-1; Medivation); MPDL3280A (PD-L1; Roche); MEDI4736 (PD-L1; AstraZeneca); MSB0010718C (PD-L1; EMD Serono); AUNP12 (PD-1; Aurigene); avelumab (PD-L1; Merck); durvalumab (PD-L1; Medimmune); IMP321, a soluble Ig fusion protein (Brignone et al., 2007, J. Immunol. 179:4202-4211); the anti-B7-H3 antibody MGA271 (Loo et al., 2012, Clin. Cancer Res. July 15 (18) 3834); TIM3 (T-cell immunoglobulin domain and mucin domain 3) inhibitors (Fourcade et al., 2010, J. Exp. Med. 207:2175-86 and Sakuishi et al., 2010, J. Exp. Med. 207:2187-94); anti-CTLA-4 antibodies described in U.S. Pat. Nos. 5,811,097; 5,811,097; 5,855,887; 6,051,227; 6,207,157; 6,682,736; 6,984,720; and 7,605,238; tremelimumab, (ticilimumab, CP-675,206); ipilimumab (also known as 10D1, MDX-D010); PD-1 and PD-L1 blockers described in U.S. Pat. Nos. 7,488,802; 7,943,743; 8,008,449; 8,168,757; 8,217,149, and PCT Published Patent Application Nos: WO03042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, and WO2011161699; nivolumab (MDX 1106, BMS 936558, ONO 4538); lambrolizumab (MK-3475 or SCH 900475); CT-011; AMP-224; and BMS-936559 (MDX-1105-01). The foregoing references are incorporated by reference herein in their entireties.

The term “statistically significant” or “significantly” refers to statistical significance, e.g., a rejection of the null hypothesis with a p-value of less than 0.05 and generally means a two standard deviation (2SD) or greater difference.

Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term “about.” The term “about” when used in connection with percentages can mean±1%.

As used herein the term “comprising” or “comprises” is used in reference to compositions, methods, and respective component(s) thereof, that are essential to the method or composition, yet open to the inclusion of unspecified elements, whether essential or not.

The term “consisting of” refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.

As used herein the term “consisting essentially of” refers to those elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment.

The singular terms “a,” “an,” and “the” include plural referents unless context clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The abbreviation, “e.g.” is derived from the Latin exempli gratia and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.”

Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims. Definitions of common terms in immunology and molecular biology can be found in The Merck Manual of Diagnosis and Therapy, 19th Edition, published by Merck Sharp & Dohme Corp., 2011 (ISBN 978-0-911910-19-3); Robert S. Porter et al. (eds.), The Encyclopedia of Molecular Cell Biology and Molecular Medicine, published by Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology by Werner Luttmann, published by Elsevier, 2006; Janeway's Immunobiology, Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), Taylor & Francis Limited, 2014 (ISBN 0815345305, 9780815345305); Lewin's Genes XI, published by Jones & Bartlett Publishers, 2014 (ISBN-1449659055); Michael Richard Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012) (ISBN 1936113414); Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch (ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley and Sons, 2014 (ISBN 047150338X, 9780471503385), Current Protocols in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and Sons, Inc., 2005; and Current Protocols in Immunology (CPI) (John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach, Warren Strobe, (eds.) John Wiley and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737), the contents of which are all incorporated by reference herein in their entireties.

Other terms are defined herein within the description of the various aspects of the invention.

All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

Specific elements of any of the foregoing embodiments can be combined or substituted for elements in other embodiments. Furthermore, while advantages associated with certain embodiments of the disclosure have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure.

All patents and other publications identified in the specification and examples are expressly incorporated herein by reference for all purposes. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the claims which follow. Further, to the extent not already indicated, it will be understood by those of ordinary skill in the art that any one of the various embodiments herein described and illustrated can be further modified to incorporate features shown in any of the other embodiments disclosed herein.

Some embodiments of the technology described herein can be defined according to any of the following numbered paragraphs:

-   -   1. A composition, comprising:         -   a. a TCR recognition domain; and         -   one or both of:         -   b. an intracellular signaling domain; and         -   c. a first-type protein interaction domain.     -   2. A composition comprising:         -   a. a first polypeptide comprising a TCR recognition domain             and a first-type protein interaction domain; and         -   b. a signaling polypeptide comprising a second-type protein             interaction domain and an intracellular signaling domain;     -    wherein the first-type and second-type protein interaction         domains bind specifically to each other.     -   3. A composition comprising:         -   a. a first polypeptide comprising a TCR recognition domain             and a first-type protein interaction domain; and         -   b. a recognition polypeptide comprising a second recognition             domain and a third-type protein interaction domain;     -    wherein the first-type and third-type protein interaction         domains bind specifically to each other.     -   4. A composition comprising:         -   a. a first polypeptide comprising a TCR recognition domain             and a first-type protein interaction domain; and         -   b. a signaling polypeptide comprising a second-type protein             interaction domain and an intracellular signaling domain;             and         -   c. a recognition polypeptide comprising a second recognition             domain and a third-type protein interaction domain;     -    wherein the second-type and third-type protein interaction         domains compete for binding to the first-type protein         interaction domain.     -   5. The composition of any of paragraphs 3-4, wherein the         third-type protein interaction domain and first-type protein         interaction domain have a higher affinity for each other than         the second-type protein interaction domain and first-type         protein interaction domain.     -   6. A composition comprising:         -   a. a first polypeptide comprising a TCR recognition domain             and a first-type protein interaction domain;         -   b. a signaling polypeptide comprising a second-type protein             interaction domain, a fourth-type protein interaction             domain, and an intracellular signaling domain; and         -   c. a recognition polypeptide comprising a second recognition             domain and a fifth-type protein interaction domain;     -    wherein the first-type protein interaction domain and the         second-type protein interaction domain bind specifically to each         other; and     -    wherein the fourth-type protein interaction domain and the         fifth-type protein interaction domain bind specifically to each         other.     -   7. The composition of paragraph 6, wherein the fourth-type         protein interaction domain and fifth-type protein interaction         domain have a weaker affinity than the second-type protein         interaction domain and first-type protein interaction domain.     -   8. The composition of any of paragraphs 6-7, wherein the first         polypeptide further comprises a sixth-type protein interaction         domain and the recognition polypeptide further comprises a         seventh-type protein interaction domain which bind specifically         to each other.     -   9. The composition of any of paragraphs 3-8, wherein the second         recognition domain is specific for a target that is not         recognized by the TCR recognition domain.     -   10. The composition of any of paragraphs 3-9, wherein the second         recognition domain is specific for a target that is found on a         healthy and/or non-target cell and not on a diseased and/or         target cell.     -   11. The composition of any of the preceding paragraphs, wherein         the TCR recognition domain comprises a MHC (Major         Histocompatibility Complex); a MHC-peptide complex; or a         MHC-peptide fusion.     -   12. The composition of paragraph 11, wherein the peptide is a         human or non-human peptide.     -   13. The composition of any of paragraphs 11-12, wherein the         peptide is a Minor Histocompatibility Antigen (MiHA).     -   14. The composition of any of paragraphs 11-13, wherein the MHC         is a monomer, dimer, trimer, tetramer, pentamer, dextramer or         other oligomer form.     -   15. The composition of any of the preceding paragraphs, wherein         the protein interaction domains are found on an extracellular         portion of the respective polypeptides.     -   16. The composition of any of the preceding paragraphs,         -   a. wherein the protein interaction domain(s) is a leucine             zipper, or any binding pair of protein interaction domains             are collectively a pair of leucine zippers;         -   b. wherein the protein interaction domain(s) is a BZip (RR)             and/or a AZip (EE), or any binding pair of protein             interaction domains are collectively a BZip (RR) and a AZip             (EE);         -   c. wherein the protein interaction domain(s) is a             PSD95-D1g1-zo-1 (PDZ) domain;         -   d. wherein the protein interaction domain(s) is a             streptavidin and/or a streptavidin binding protein (SBP) or             any binding pair of protein interaction domains are             collectively a streptavidin and a streptavidin binding             protein (SBP);         -   e. wherein the protein interaction domain(s) is a             FKBP-binding domain of mTOR (FRB) and/or a FK506 binding             protein (FKBP) or any binding pair of protein interaction             domains are collectively a FKBP-binding domain of mTOR (FRB)             and a FK506 binding protein (FKBP);         -   f. wherein the protein interaction domain(s) is a             cyclophilin-Fas fusion protein (CyP-Fas) and/or a FK506             binding protein (FKBP) or any binding pair of protein             interaction domains are collectively a cyclophilin-Fas             fusion protein (CyP-Fas) and a FK506 binding protein (FKBP);         -   g. wherein the protein interaction domain(s) is a             calcineurinA (CNA) and/or a FK506 binding protein (FKBP) or             any binding pair of protein interaction domains are             collectively a calcineurinA (CNA) and a FK506 binding             protein (FKBP);         -   h. wherein the protein interaction domain(s) is a             gibberellin insensitive (GIA) and/or a gibberellin             insensitive dwarf1 (GID1) or any binding pair of protein             interaction domains are collectively a gibberellin             insensitive (GIA) and a gibberellin insensitive dwarf1             (GID1);         -   i. wherein the protein interaction domain(s) is a Snap-tag             and/or a Halo tag, or any binding pair of protein             interaction domains are collectively a Snap-tag and a Halo             tag;         -   j. wherein the protein interaction domain(s) is a             T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52),             or any binding pair of protein interaction domains are             collectively a T14-3-3-cdeltaC and a C-Terminal peptides of             PMA2 (CT52);         -   k. wherein the protein interaction domain(s) is a PYL and/or             a ABI, or any binding pair of protein interaction domains             are collectively a PYL and a ABI; and/or         -   l. wherein the protein interaction domain(s) is a nucleotide             tag and/or a zinc finger domain, or any binding pair of             protein interaction domains are collectively a nucleotide             tag and a zinc finger domain.     -   17. The composition of paragraph 16, wherein the nucleotide tag         is a DNA tag or dsDNA tag.     -   18. The composition of any of the preceding paragraphs, wherein         the intracellular signaling domain is a signaling domain from a         protein selected from the group consisting of:         -   TCRC; FcRy; FcRp; CD3y; CD35; CD3s; CD3C; CD22; CD79a;             CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54             (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1);             CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2);             CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM;             ZAP70; and 41BB.     -   19. A cell comprising and/or expressing the composition of any         of the preceding paragraphs.     -   20. The cell of paragraph 19, wherein the TCR recognition domain         comprises a MHC allogenic to the cell.     -   21. The cell of paragraph 19, wherein the TCR recognition domain         comprises a peptide allogenic to the cell.     -   22. The cell of any of paragraphs 19-21, wherein the cell is a         dendritic cell, regulatory T cell, or effector T cell.     -   23. The cell of any of paragraphs 19-22, wherein the cell is         engineered to express the polypeptide(s) of the composition.     -   24. The cell of any of paragraphs 19-22, wherein the cell is         engineered to express the signaling polypeptide of the         composition.     -   25. The cell of any of paragraphs 19-24, wherein the cell is         further engineered to knockout the native MHCI/II.     -   26. A chimeric antigen receptor (CAR) comprising:         -   a. an anti-CD127 and/or anti-CD45RO recognition domain;         -   b. an intracellular signaling domain.     -   27. A composition comprising:         -   a first polypeptide comprising:         -   a. an anti-CD127 and/or anti-CD45RO recognition domain;         -   b. a first-type protein interaction domain; and         -   a second polypeptide comprising:         -   c. a second-type protein interaction domain; and         -   d. an intracellular signaling domain;     -    wherein the first-type protein interaction domain and the         second-type protein interaction domain bind specifically to each         other.     -   28. A composition comprising:         -   a first polypeptide comprising:         -   a. an anti-CD127 recognition domain;         -   b. a first-type protein interaction domain;         -   a second polypeptide comprising:         -   c. an anti-CD45RO recognition domain;         -   d. a fifth-type protein interaction domain; and         -   a third polypeptide comprising:         -   e. a second-type and a fourth-type protein interaction             domain; and         -   f. an intracellular signaling domain;     -    wherein the first-type protein interaction domain and the         second-type protein interaction domain bind specifically to each         other; and     -    wherein the fourth-type protein interaction domain and the         fifth-type protein interaction domain bind specifically to each         other.     -   29. A cell comprising the CAR or composition of any of         paragraphs 26-28.     -   30. A method of treating an autoimmune disease or treating or         preventing transplant rejection or GVHD in a subject in need         thereof, the method comprising administering to the subject a         composition and/or cell of any of the preceding paragraphs.     -   31. The method of paragraph 30, wherein the TCR recognition         domain comprises a MHC allogenic to the subject.     -   32. The method of paragraph 30, wherein the TCR recognition         domain comprises a MHC autologous to the transplant cells.     -   33. The method of paragraph 30, wherein the TCR recognition         domain comprises a peptide allogenic to the subject.     -   34. The method of paragraph 30, wherein the TCR recognition         domain comprises a peptide autologous to the transplant cells.     -   35. The method of any of paragraphs 32 or 34, wherein the         transplant is vascularized composite allotransplantation (VCA).     -   36. The method of any of paragraphs 35, wherein the autoimmune         disease is type 1 diabetes, multiple sclerosis, rheumatoid         arthritis, or scleroderma.

Some embodiments of the technology described herein can be defined according to any of the following numbered paragraphs:

-   -   1. A composition, comprising:         -   a. a TCR recognition domain; and         -   one or both of:         -   b. an intracellular signaling domain; and         -   c. a first-type biomolecular interaction domain.     -   2. A composition comprising:         -   a. a first polypeptide comprising at least a portion of a             TCR recognition domain and a first-type biomolecular             interaction domain; and         -   b. a signaling polypeptide comprising a second-type             biomolecular interaction domain and an intracellular             signaling domain;     -    wherein the first-type and second-type biomolecular interaction         domains bind specifically to each other.     -   3. A composition comprising:         -   a. a first polypeptide comprising at least a portion of a             TCR recognition domain and a first-type biomolecular             interaction domain; and         -   b. a recognition polypeptide comprising a second recognition             domain and a third-type biomolecular interaction domain;     -    wherein the first-type and third-type biomolecular interaction         domains bind specifically to each other.     -   4. A composition comprising:         -   a. a first polypeptide comprising at least a portion of a             TCR recognition domain and a first-type biomolecular             interaction domain; and         -   b. a signaling polypeptide comprising a second-type             biomolecular interaction domain and an intracellular             signaling domain; and         -   c. a recognition polypeptide comprising a second recognition             domain and a third-type biomolecular interaction domain;     -    wherein the second-type and third-type biomolecular interaction         domains compete for binding to the first-type biomolecular         interaction domain.     -   5. The composition of any of paragraphs 3-4, wherein the         third-type biomolecular interaction domain and first-type         biomolecular interaction domain have a higher affinity for each         other than the second-type biomolecular interaction domain and         first-type biomolecular interaction domain.     -   6. A composition comprising:         -   a. a first polypeptide comprising at least a portion of a             TCR recognition domain and a first-type biomolecular             interaction domain;         -   b. a signaling polypeptide comprising a second-type             biomolecular interaction domain, a fourth-type biomolecular             interaction domain, and an intracellular signaling domain;             and         -   c. a recognition polypeptide comprising a second recognition             domain and a fifth-type biomolecular interaction domain;     -    wherein the first-type biomolecular interaction domain and the         second-type biomolecular interaction domain bind specifically to         each other; and     -    wherein the fourth-type biomolecular interaction domain and the         fifth-type biomolecular interaction domain bind specifically to         each other.     -   7. The composition of paragraph 6, wherein the fourth-type         biomolecular interaction domain and fifth-type biomolecular         interaction domain have a weaker affinity than the second-type         biomolecular interaction domain and first-type protein         interaction domain.     -   8. The composition of any of paragraphs 6-7, wherein the first         polypeptide further comprises a sixth-type biomolecular         interaction domain and the recognition polypeptide further         comprises a seventh-type biomolecular interaction domain which         bind specifically to each other.     -   9. The composition of any of paragraphs 2-8, wherein the first         polypeptide comprises the entire TCR recognition domain.     -   10. The composition of any of paragraphs 2-8, wherein the TCR         recognition domain comprises at least two separate polypeptide         sequences, the first polypeptide comprises at least one of the         separate polypeptide sequences of the TCR recognition domain,         and the first polypeptide is bound to or complexed with a second         or further polypeptide sequences of the TCR recognition domain         to form a TCR recognition domain.     -   11. The composition of any of paragraphs 1-10, wherein the TCR         recognition domain comprises a non-polypeptide component.     -   12. The composition of any of paragraphs 3-11, wherein the         second recognition domain is specific for a target that is not         recognized by the TCR recognition domain.     -   13. The composition of any of paragraphs 3-12, wherein the         second recognition domain is specific for a target that is found         on a healthy and/or non-target cell and not on a diseased and/or         target cell.     -   14. The composition of any of the preceding paragraphs, wherein         the TCR recognition domain comprises a MHC (Major         Histocompatibility Complex); a MHC-peptide complex; featureless         peptide MHC; or a MHC-peptide fusion.     -   15. The composition of paragraph 14, wherein the peptide is a         human or non-human peptide.     -   16. The composition of any of paragraphs 14-15, wherein the         peptide is a Minor Histocompatibility     -   Antigen (MiHA).     -   17. The composition of any of paragraphs 14-16, wherein the MHC         is a monomer, dimer, trimer, tetramer, pentamer, dextramer or         other oligomer form.     -   18. The composition of any of paragraphs 14-17, wherein the         MHC-peptide complex is a monomer, dimer, trimer, tetramer,         pentamer, dextramer or other oligomer form.     -   19. The composition of any of paragraphs 14-17, wherein the         MHC-peptide fusion is a monomer, dimer, trimer, tetramer,         pentamer, dextramer or other oligomer form.     -   20. The composition of any of paragraphs 14-17, wherein the MHC         is a dimer, trimer, tetramer, pentamer, dextramer or other         oligomer form.     -   21. The composition of any of paragraphs 14-17, wherein the         MHC-peptide complex is a dimer, trimer, tetramer, pentamer,         dextramer or other oligomer form.     -   22. The composition of any of paragraphs 14-17, wherein the         MHC-peptide fusion is a dimer, trimer, tetramer, pentamer,         dextramer or other oligomer form.     -   23. The composition of any of paragraphs 14-22, wherein the MHC         is a MHC class I or a MHC class II.     -   24. The composition of any of paragraphs 1-13, wherein the TCR         recognition domain comprises a CD1 domain or a CD1 domain-ligand         complex or fusion.     -   25. The composition of paragraph 24, wherein the CD1 is CD1d.     -   26. The composition of any of the preceding paragraphs, wherein         the biomolecular interaction domains are found on an         extracellular portion of the respective polypeptides.     -   27. The composition of any of the preceding paragraphs,         -   a. wherein the biomolecular interaction domain(s) is a             leucine zipper, or any binding pair of biomolecular             interaction domains are collectively a pair of leucine             zippers;         -   b. wherein the biomolecular interaction domain(s) is a BZip             (RR) and/or a AZip (EE), or any binding pair of biomolecular             interaction domains are collectively a BZip (RR) and a AZip             (EE);         -   c. wherein the biomolecular interaction domain(s) is a             PSD95-D1g1-zo-1 (PDZ) domain;         -   d. wherein the biomolecular interaction domain(s) is a             streptavidin and/or a streptavidin binding biomolecular             (SBP) or any binding pair of biomolecular interaction             domains are collectively a streptavidin and a streptavidin             binding biomolecular (SBP);         -   e. wherein the biomolecular interaction domain(s) is a             FKBP-binding domain of mTOR (FRB) and/or a FK506 binding             biomolecular (FKBP) or any binding pair of biomolecular             interaction domains are collectively a FKBP-binding domain             of mTOR (FRB) and a FK506 binding biomolecular (FKBP);         -   f. wherein the biomolecular interaction domain(s) is a             cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506             binding biomolecular (FKBP) or any binding pair of             biomolecular interaction domains are collectively a             cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506             binding biomolecular (FKBP);         -   g. wherein the biomolecular interaction domain(s) is a             calcineurin A (CNA) and/or a FK506 binding biomolecular             (FKBP) or any binding pair of biomolecular interaction             domains are collectively a calcineurin A (CNA) and a FK506             binding biomolecular (FKBP);         -   h. wherein the biomolecular interaction domain(s) is a             gibberellin insensitive (GIA) and/or a gibberellin             insensitive dwarf1 (GID1) or any binding pair of             biomolecular interaction domains are collectively a             gibberellin insensitive (GIA) and a gibberellin insensitive             dwarf1 (GID1);         -   i. wherein the biomolecular interaction domain(s) is a             Snap-tag and/or a Halo tag, or any binding pair of             biomolecular interaction domains are collectively a Snap-tag             and a Halo tag;         -   j. wherein the biomolecular interaction domain(s) is a             T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52),             or any binding pair of biomolecular interaction domains are             collectively a T14-3-3-cdeltaC and a C-Terminal peptides of             PMA2 (CT52);         -   k. wherein the biomolecular interaction domain(s) is a PYL             and/or a ABI, or any binding pair of biomolecular             interaction domains are collectively a PYL and a ABI;         -   l. wherein the biomolecular interaction domain(s) is a             nucleotide tag and/or a zinc finger domain, or any binding             pair of biomolecular interaction domains are collectively a             nucleotide tag and a zinc finger domain;         -   m. wherein the biomolecular interaction domain(s) is a             nucleotide tag, or any binding pair of biomolecular             interaction domains are collectively a pair of nucleotide             tags;         -   n. wherein the biomolecular interaction domain(s) is a             Fluorescein isothiocyanate (FITC) and/or a FITC binding             biomolecular or any binding pair of protein interaction             domains are collectively a FITC and a FITC binding protein;             and/or         -   o. wherein the protein interaction domain(s) is a             (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein             or any binding pair of protein interaction domains are             collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE             binding protein.     -   28. The composition of paragraph 27, wherein the nucleotide tag         is a DNA tag or dsDNA tag.     -   29. The composition of any of the preceding paragraphs, wherein         the intracellular signaling domain comprises or is a signaling         domain from one or more proteins selected from the group         consisting of:         -   TCRζ, FcRγ, FcRβ, CD3γ; CD35; CD3ζ; CD3C; CD22; CD79a;             CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54             (ICAM); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1);             CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2);             CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM;             and ZAP70.     -   30. A cell comprising and/or expressing the composition of any         of the preceding paragraphs.     -   31. A composition comprising a first polypeptide of any of the         preceding paragraphs and a cell expressing or comprising the         signaling polypeptide of any of the preceding paragraphs.     -   32. The cell or composition of any of paragraphs 30-31, wherein         the TCR recognition domain comprises a MHC allogeneic,         autologous, or xenogeneic to the cell.     -   33. The cell or composition of any of paragraphs 30-32, wherein         the TCR recognition domain comprises a synthetic MHC.     -   34. The cell or composition of any of paragraphs 30-33, wherein         the TCR recognition domain comprises a MHC and a peptide,         wherein the peptide is allogeneic, autologous, or xenogeneic to         the cell.     -   35. The cell or composition of any of paragraphs 30-34, wherein         the TCR recognition domain comprises a MHC and a peptide,         wherein the peptide is synthetic.     -   36. The cell or composition of any of paragraphs 30-35, wherein         the cell is a NK cell, dendritic cell, regulatory T cell, or         effector T cell.     -   37. The cell or composition of any of paragraphs 30-36, wherein         the cell is engineered to express one of more of the         polypeptide(s) of the composition.     -   38. The cell or composition of any of paragraphs 30-37, wherein         the cell is engineered to express the signaling polypeptide of         the composition.     -   39. The cell or composition of any of paragraphs 30-38, wherein         the cell is further engineered to knockout or knockdown the         native MHCI/II.     -   40. The cell or composition of any of paragraphs 30-39, wherein         the cell is further engineered to knockdown the native MHCI/II         expressed on the cell surface.     -   41. A chimeric antigen receptor (CAR) comprising:         -   a. an anti-CD127 and/or anti-CD45RO recognition domain;         -   b. an intracellular signaling domain.     -   42. A composition comprising:         -   a first polypeptide comprising:         -   a. an anti-CD127 and/or anti-CD45RO recognition domain;         -   b. a first-type protein interaction domain; and         -   a second polypeptide comprising:         -   c. a second-type protein interaction domain; and         -   d. an intracellular signaling domain;     -    wherein the first-type protein interaction domain and the         second-type protein interaction domain bind specifically to each         other.     -   43. A composition comprising:         -   a first polypeptide comprising:         -   a. an anti-CD127 recognition domain;         -   b. a first-type protein interaction domain;         -   a second polypeptide comprising:         -   c. an anti-CD45RO recognition domain;         -   d. a fifth-type protein interaction domain; and         -   a third polypeptide comprising:         -   e. a second-type and a fourth-type protein interaction             domain; and         -   f. an intracellular signaling domain;     -    wherein the first-type protein interaction domain and the         second-type protein interaction domain bind specifically to each         other; and     -    wherein the fourth-type protein interaction domain and the         fifth-type protein interaction domain bind specifically to each         other.     -   44. A cell comprising the CAR or composition of any of         paragraphs 41-43.     -   45. A method of treating or preventing an autoimmune disease or         condition; T cell mediated inflammation or immune response;         malignant T cell condition; transplant rejection; or GvHD in a         subject in need thereof, the method comprising administering to         the subject a composition and/or cell of any of the preceding         paragraphs.     -   46. The method of paragraph 45, wherein the TCR recognition         domain comprises a MHC allogeneic to the subject.     -   47. The method of paragraph 45, wherein the TCR recognition         domain comprises a MHC autologous to the transplant cells.     -   48. The method of paragraph 45, wherein the TCR recognition         domain comprises a MHC xenogeneic to the transplant cells.     -   49. The method of paragraph 45, wherein the TCR recognition         domain comprises a MHC and a peptide, wherein the peptide is         allogeneic to the subject.     -   50. The method of paragraph 45, wherein the TCR recognition         domain comprises a MHC and a peptide, wherein the peptide is         autologous to the transplant cells.     -   51. The method of paragraph 45, wherein the TCR recognition         domain comprises a MHC and a peptide, wherein the peptide is         autologous to the transplant cells.     -   52. The method of any of paragraphs 45-51, wherein the MHC         and/or the peptide is synthetic.     -   53. The method of any of paragraphs 45-52, wherein the         transplant is any human or non-human cell, tissue, or organ.     -   54. The method of any of paragraphs 45-53, wherein the         transplant is an allogeneic hematopoietic stem cell or solid         organ transplantation.     -   55. The method of any of paragraphs 45-52, wherein the malignant         T cell condition is T cell acute lymphoblastic leukemia or T         cell lymphoblastic lymphoma.     -   56. The method of any of paragraphs 45-52, wherein the         autoimmune disease is type 1 diabetes, vitiligo, multiple         sclerosis, alopecia, celiac disease, pemphigus, rheumatoid         arthritis, or scleroderma.     -   57. The method of any of paragraphs 45-52, wherein the         autoimmune disease is thyroiditis, type 1 diabetes mellitus,         Hashimoto's thyroiditis, Graves' disease, celiac disease,         multiple sclerosis, Guillain-Barre syndrome, Addison's disease,         and Raynaud's phenomenon, Goodpasture's disease, arthritis         (rheumatoid arthritis such as acute arthritis, chronic         rheumatoid arthritis, gout or gouty arthritis, acute gouty         arthritis, acute immunological arthritis, chronic inflammatory         arthritis, degenerative arthritis, type II collagen-induced         arthritis, infectious arthritis, Lyme arthritis (e.g., post         treatment Lyme disease syndrome), proliferative arthritis,         psoriatic arthritis, Still's disease, vertebral arthritis, and         juvenile-onset rheumatoid arthritis, arthritis chronica         progrediente, arthritis deformans, polyarthritis chronica         primaria, reactive arthritis, and ankylosing spondylitis),         palindromic arthritis, inflammatory hyperproliferative skin         diseases, psoriasis such as plaque psoriasis, guttate psoriasis,         pustular psoriasis, and psoriasis of the nails, atopy including         atopic diseases such as hay fever and Job's syndrome, dermatitis         including contact dermatitis, chronic contact dermatitis,         exfoliative dermatitis, allergic dermatitis, allergic contact         dermatitis, dermatitis herpetiformis, nummular dermatitis,         seborrheic dermatitis, non-specific dermatitis, primary irritant         contact dermatitis, and atopic dermatitis, x-linked hyper IgM         syndrome, allergic intraocular inflammatory diseases, urticaria         such as chronic allergic urticaria and chronic idiopathic         urticaria, including chronic autoimmune urticaria, myositis,         polymyositis/dermatomyositis, juvenile dermatomyositis, toxic         epidermal necrolysis, scleroderma (including systemic         scleroderma), sclerosis such as systemic sclerosis, multiple         sclerosis (MS) such as spino-optical MS, primary progressive MS         (PPMS), and relapsing remitting MS (RRMS), progressive systemic         sclerosis, atherosclerosis, arteriosclerosis, sclerosis         disseminata, ataxic sclerosis, neuromyelitis optica (NMO),         inflammatory bowel disease (IBD) (for example, Crohn's disease,         autoimmune-mediated gastrointestinal diseases, colitis such as         ulcerative colitis, colitis ulcerosa, microscopic colitis,         collagenous colitis, colitis polyposa, necrotizing         enterocolitis, and transmural colitis, and autoimmune         inflammatory bowel disease), bowel inflammation, pyoderma         gangrenosum, erythema nodosum, primary sclerosing cholangitis,         respiratory distress syndrome, including adult or acute         respiratory distress syndrome (ARDS), meningitis, inflammation         of all or part of the uvea, iritis, choroiditis, an autoimmune         hematological disorder, rheumatoid spondylitis, rheumatoid         synovitis, hereditary angioedema, cranial nerve damage as in         meningitis, herpes gestationis, pemphigoid gestationis, pruritis         scroti, autoimmune premature ovarian failure, sudden hearing         loss due to an autoimmune condition, IgE-mediated diseases such         as anaphylaxis and allergic and atopic rhinitis, encephalitis         such as Rasmussen's encephalitis and limbic and/or brainstem         encephalitis, uveitis, such as anterior uveitis, acute anterior         uveitis, granulomatous uveitis, nongranulomatous uveitis,         phacoantigenic uveitis, posterior uveitis, or autoimmune         uveitis, glomerulonephritis (GN) with and without nephrotic         syndrome such as chronic or acute glomerulonephritis such as         primary GN, immune-mediated GN, membranous GN (membranous         nephropathy), idiopathic membranous GN or idiopathic membranous         nephropathy, membrano- or membranous proliferative GN (MPGN),         including Type I and Type II, and rapidly progressive GN,         proliferative nephritis, autoimmune polyglandular endocrine         failure, balanitis including balanitis circumscripta         plasmacellularis, balanoposthitis, erythema annulare         centrifugum, erythema dyschromicum perstans, eythema multiform,         granuloma annulare, lichen nitidus, lichen sclerosus et         atrophicus, lichen simplex chronicus, lichen spinulosus, lichen         planus, lamellar ichthyosis, epidermolytic hyperkeratosis,         premalignant keratosis, pyoderma gangrenosum, allergic         conditions and responses, allergic reaction, eczema including         allergic or atopic eczema, asteatotic eczema, dyshidrotic         eczema, and vesicular palmoplantar eczema, asthma such as asthma         bronchiale, bronchial asthma, and auto-immune asthma, conditions         involving infiltration of T cells and chronic inflammatory         responses, immune reactions against foreign antigens such as         fetal A-B-O blood groups during pregnancy, chronic pulmonary         inflammatory disease, autoimmune myocarditis, leukocyte adhesion         deficiency, lupus, including lupus nephritis, lupus cerebritis,         pediatric lupus, non-renal lupus, extra-renal lupus, discoid         lupus and discoid lupus erythematosus, alopecia lupus, systemic         lupus erythematosus (SLE) such as cutaneous SLE or subacute         cutaneous SLE, neonatal lupus syndrome (NLE), and lupus         erythematosus disseminatus, juvenile onset (Type I) diabetes         mellitus, including pediatric insulin-dependent diabetes         mellitus (IDDM), adult onset diabetes mellitus (Type II         diabetes), autoimmune diabetes, idiopathic diabetes insipidus,         diabetic retinopathy, diabetic nephropathy, diabetic         large-artery disorder, immune responses associated with acute         and delayed hypersensitivity mediated by cytokines and         T-lymphocytes, sarcoidosis, granulomatosis including         lymphomatoid granulomatosis, Wegener's granulomatosis,         agranulocytosis, vasculitides, including vasculitis,         large-vessel vasculitis (including polymyalgia rheumatica and         giant-cell (Takayasu's) arteritis), medium-vessel vasculitis         (including Kawasaki's disease and polyarteritis         nodosa/periarteritis nodosa), microscopic polyarteritis,         immunovasculitis, CNS vasculitis, cutaneous vasculitis,         hypersensitivity vasculitis, necrotizing vasculitis such as         systemic necrotizing vasculitis, and ANCA-associated vasculitis,         such as Churg-Strauss vasculitis or syndrome (CSS) and         ANCA-associated small-vessel vasculitis, temporal arteritis,         autoimmune aplastic anemia, Coombs positive anemia, Diamond         Blackfan anemia, hemolytic anemia or immune hemolytic anemia         including autoimmune hemolytic anemia (AIHA), pernicious anemia         (anemia perniciosa), Addison's disease, pure red cell anemia or         aplasia (PRCA), Factor VIII deficiency, hemophilia A, autoimmune         neutropenia, pancytopenia, leukopenia, diseases involving         leukocyte diapedesis, CNS inflammatory disorders, multiple organ         injury syndrome such as those secondary to septicemia, trauma or         hemorrhage, antigen-antibody complex-mediated diseases,         anti-glomerular basement membrane disease, anti-phospholipid         antibody syndrome, allergic neuritis, Behcet's disease/syndrome,         Castleman's syndrome, Goodpasture's syndrome, Reynaud's         syndrome, Sjogren's syndrome, Stevens-Johnson syndrome,         pemphigoid such as pemphigoid bullous and skin pemphigoid,         pemphigus (including pemphigus vulgaris, pemphigus foliaceus,         pemphigus mucus-membrane pemphigoid, and pemphigus         erythematosus), autoimmune polyendocrinopathies, Reiter's         disease or syndrome, an immune complex disorder such as immune         complex nephritis, antibody-mediated nephritis,         polyneuropathies, chronic neuropathy such as IgM         polyneuropathies or IgM-mediated neuropathy, and autoimmune or         immune-mediated thrombocytopenia such as idiopathic         thrombocytopenic purpura (ITP) including chronic or acute ITP,         scleritis such as idiopathic cerato-scleritis, episcleritis,         autoimmune disease of the testis and ovary including autoimmune         orchitis and oophoritis, primary hypothyroidism,         hypoparathyroidism, autoimmune endocrine diseases including         thyroiditis such as autoimmune thyroiditis, Hashimoto's disease,         chronic thyroiditis (Hashimoto's thyroiditis), or subacute         thyroiditis, idiopathic hypothyroidism, Grave's disease,         polyglandular syndromes such as autoimmune polyglandular         syndromes (or polyglandular endocrinopathy syndromes),         paraneoplastic syndromes, including neurologic paraneoplastic         syndromes such as Lambert-Eaton myasthenic syndrome or         Eaton-Lambert syndrome, stiff-man or stiff-person syndrome,         encephalomyelitis such as allergic encephalomyelitis or         encephalomyelitis allergica and experimental allergic         encephalomyelitis (EAE), myasthenia gravis such as         thymoma-associated myasthenia gravis, cerebellar degeneration,         neuromyotonia, opsoclonus or opsoclonus myoclonus syndrome         (OMS), and sensory neuropathy, multifocal motor neuropathy,         Sheehan's syndrome, autoimmune hepatitis, lupoid hepatitis,         giant-cell hepatitis, autoimmune chronic active hepatitis,         lymphoid interstitial pneumonitis (LIP), bronchiolitis         obliterans (non-transplant) vs NSIP, Guillain-Barre syndrome,         Berger's disease (IgA nephropathy), idiopathic IgA nephropathy,         linear IgA dermatosis, acute febrile neutrophilic dermatosis,         subcorneal pustular dermatosis, transient acantholytic         dermatosis, cirrhosis such as primary biliary cirrhosis and         pneumonocirrhosis, autoimmune enteropathy syndrome, Celiac or         Coeliac disease, celiac sprue (gluten enteropathy), refractory         sprue, idiopathic sprue, cryoglobulinemia, amylotrophic lateral         sclerosis (ALS; Lou Gehrig's disease), coronary artery disease,         autoimmune ear disease such as autoimmune inner ear disease         (AIED), autoimmune hearing loss, polychondritis such as         refractory or relapsed or relapsing polychondritis, pulmonary         alveolar proteinosis, Cogan's syndrome/nonsyphilitic         interstitial keratitis, Bell's palsy, Sweet's disease/syndrome,         rosacea autoimmune, zoster-associated pain, amyloidosis, a         non-cancerous lymphocytosis, a primary lymphocytosis, which         includes monoclonal B cell lymphocytosis (e.g., benign         monoclonal gammopathy and monoclonal gammopathy of undetermined         significance, MGUS), peripheral neuropathy, paraneoplastic         syndrome, channelopathies including channelopathies of the CNS,         autism, inflammatory myopathy, focal or segmental or focal         segmental glomerulosclerosis (FSGS), endocrine opthalmopathy,         uveoretinitis, chorioretinitis, autoimmune hepatological         disorder, fibromyalgia, multiple endocrine failure, Schmidt's         syndrome, adrenalitis, gastric atrophy, presenile dementia,         demyelinating diseases such as autoimmune demyelinating diseases         and chronic inflammatory demyelinating polyneuropathy,         Dressler's syndrome, alopecia areata, alopecia totalis, CREST         syndrome (calcinosis, Raynaud's phenomenon, esophageal         dysmotility, sclerodactyly, and telangiectasia), male and female         autoimmune infertility, e.g., due to anti-spermatozoan         antibodies, mixed connective tissue disease, Chagas' disease,         rheumatic fever, recurrent abortion, farmer's lung, erythema         multiforme, post-cardiotomy syndrome, Cushing's syndrome,         bird-fancier's lung, allergic granulomatous angiitis, benign         lymphocytic angiitis, Alport's syndrome, alveolitis such as         allergic alveolitis and fibrosing alveolitis, interstitial lung         disease, transfusion reaction, Sampter's syndrome, Caplan's         syndrome, endocarditis, endomyocardial fibrosis, diffuse         interstitial pulmonary fibrosis, interstitial lung fibrosis,         pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic         fibrosis, endophthalmitis, erythema elevatum et diutinum,         erythroblastosis fetalis, eosinophilic faciitis, Shulman's         syndrome, Felty's syndrome, cyclitis such as chronic cyclitis,         heterochronic cyclitis, iridocyclitis (acute or chronic), or         Fuch's cyclitis, Henoch-Schonlein purpura, SCID, sepsis,         endotoxemia, post-vaccination syndromes, Evan's syndrome,         autoimmune gonadal failure, Sydenham's chorea,         post-streptococcal nephritis, thromboangitis ubiterans,         thyrotoxicosis, tabes dorsalis, chorioiditis, giant-cell         polymyalgia, chronic hypersensitivity pneumonitis,         keratoconjunctivitis sicca, idiopathic nephritic syndrome,         minimal change nephropathy, benign familial and         ischemia-reperfusion injury, transplant organ reperfusion,         retinal autoimmunity, aphthae, aphthous stomatitis,         arteriosclerotic disorders, aspermiogenesis, autoimmune         hemolysis, Boeck's disease, enteritis allergica, erythema         nodosum leprosum, idiopathic facial paralysis, chronic fatigue         syndrome, febris rheumatica, Hamman-Rich's disease, sensoneural         hearing loss, ileitis regionalis, leucopenia, transverse         myelitis, primary idiopathic myxedema, ophthalmia symphatica,         polyradiculitis acuta, pyoderma gangrenosum, acquired spenic         atrophy, vitiligo, toxic-shock syndrome, conditions involving         infiltration of T cells, leukocyte-adhesion deficiency, immune         responses associated with acute and delayed hypersensitivity         mediated by cytokines and T-lymphocytes, diseases involving         leukocyte diapedesis, multiple organ injury syndrome,         antigen-antibody complex-mediated diseases, antiglomerular         basement membrane disease, allergic neuritis, autoimmune         polyendocrinopathies, oophoritis, primary myxedema, autoimmune         atrophic gastritis, rheumatic diseases, mixed connective tissue         disease, nephrotic syndrome, insulitis, polyendocrine failure,         autoimmune polyglandular syndrome type I, adult-onset idiopathic         hypoparathyroidism (AOIH), myocarditis, nephrotic syndrome,         primary sclerosing cholangitis, acute or chronic sinusitis,         ethmoid, frontal, maxillary, or sphenoid sinusitis, an         eosinophil-related disorder such as eosinophilia, pulmonary         infiltration eosinophilia, eosinophilia-myalgia syndrome,         Loffler's syndrome, chronic eosinophilic pneumonia, tropical         pulmonary eosinophilia, granulomas containing eosinophils,         seronegative spondyloarthritides, polyendocrine autoimmune         disease, sclerosing cholangitis, sclera, episclera, Bruton's         syndrome, transient hypogammaglobulinemia of infancy,         Wiskott-Aldrich syndrome, ataxia telangiectasia syndrome,         angiectasis, autoimmune disorders associated with collagen         disease, rheumatism, allergic hypersensitivity disorders,         glomerulonephritides, reperfusion injury, ischemic re-perfusion         disorder, lymphomatous tracheobronchitis, inflammatory         dermatoses, dermatoses with acute inflammatory components, and         autoimmune uveoretinitis (AUR).     -   58. The method of any of paragraphs 45-52, wherein the T cell         mediated immune response is an anti-drug specific response to a         biologic, cell therapy, and/or gene therapy.     -   59. The method of paragraph 58, wherein the biologic,         cell-therapy, or gene therapy is an adeno-associated virus (AAV)         gene therapy, a genome editing agent, or enzyme replacement         therapy.     -   60. The method of any of paragraphs 45-52, wherein the disease         is type 1 diabetes and the TCR recognition domain comprises         sequences with at least 80% or at least 95% sequence identity         to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one         of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at         least one of SEQ ID NOs: 20128-2129.     -   61. The method of any of paragraphs 45-52, wherein the disease         is vitiligo and the TCR recognition domain comprises sequences         with at least 80% or at least 95% sequence identity to: SEQ ID         NO: 18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID         NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises         HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO:         2021.     -   62. The method of any of paragraphs 45-52, wherein the method is         a method of treating and/or preventing GvHD and the TCR         recognition domain comprises sequences with at least 80% or at         least 95% sequence identity to: HLA-A*0101 and at least one of         SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or         HLA-B*0801 and SEQ ID NO: 2039.     -   63. The method of any of paragraphs 45-52, wherein the disease         is type 1 diabetes and the TCR recognition domain comprises one         or more of SEQ ID NOs: 8-17; comprises HLA-A*0201 and at least         one of SEQ ID NOs: 2013-2016 and 2031-2033; or comprises         HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.     -   64. The method of any of paragraphs 45-52, wherein the disease         is vitiligo and the TCR recognition domain comprises SEQ ID NO:         18, 19, and one of 20-22; or comprises HLA-A*0201 and SEQ ID NO:         2018; or comprises HLA-A*0301 and SEQ ID NO: 2019; or comprises         HLA-A*2402 and SEQ ID NO: 2020; or comprises HLA-A*0101 and SEQ         ID NO: 2021.     -   65. The method of any of paragraphs 45-52, wherein the method is         a method of treating and/or preventing GvHD and the TCR         recognition domain comprises HLA-A*0101 and at least one of SEQ         ID NOs: 2034-2037; or comprises HLA-B*0702 and SEQ ID NO: 2038;         or comprises HLA-B*0801 and SEQ ID NO: 2039.

Some embodiments of the technology described herein can be defined according to any of the following numbered paragraphs:

-   -   1. A composition, comprising:         -   a. a TCR recognition domain; and         -   one or both of:         -   b. an intracellular signaling domain; and         -   c. a biomolecular interaction domain.     -   2. The composition of paragraph 1, comprising a TCR recognition         domain and a biomolecular interaction domain.     -   3. The composition of paragraph 1, comprising a TCR recognition         domain and an intracellular signaling domain.     -   4. The composition of paragraph 1, wherein the biomolecular         interaction domain of c) is a first-type biomolecular         interaction domain and the composition further comprises a         signaling polypeptide comprising a second-type biomolecular         interaction domain and an intracellular signaling domain;         wherein the first-type and second-type biomolecular interaction         domains bind specifically to each other.     -   5. The composition of paragraph 1, wherein the TCR recognition         domain comprises a MHC (Major Histocompatibility Complex); a         MHC-peptide complex; a featureless peptide MHC; or a MHC-peptide         fusion.     -   6. The composition of paragraph 1, wherein the TCR recognition         domain comprises a CD1 domain or a CD1 domain-ligand complex or         fusion.     -   7. The composition of paragraph 6, wherein the CD1 is CD1d.     -   8. The composition of paragraph 1, wherein the TCR recognition         domain is a monomer, dimer, trimer, tetramer, pentamer,         dextramer or other oligomer form.     -   9. The composition of paragraph 1,         -   a. wherein the biomolecular interaction domain(s) is a             leucine zipper, or any binding pair of biomolecular             interaction domains are collectively a pair of leucine             zippers;         -   b. wherein the biomolecular interaction domain(s) is a BZip             (RR) and/or a AZip (EE), or any binding pair of biomolecular             interaction domains are collectively a BZip (RR) and a AZip             (EE);         -   c. wherein the biomolecular interaction domain(s) is a             PSD95-D1g1-zo-1 (PDZ) domain;         -   d. wherein the biomolecular interaction domain(s) is a             streptavidin and/or a streptavidin binding biomolecular             (SBP) or any binding pair of biomolecular interaction             domains are collectively a streptavidin and a streptavidin             binding biomolecular (SBP);         -   e. wherein the biomolecular interaction domain(s) is a             FKBP-binding domain of mTOR (FRB) and/or a FK506 binding             biomolecular (FKBP) or any binding pair of biomolecular             interaction domains are collectively a FKBP-binding domain             of mTOR (FRB) and a FK506 binding biomolecular (FKBP);         -   f. wherein the biomolecular interaction domain(s) is a             cyclophilin-Fas fusion biomolecular (CyP-Fas) and/or a FK506             binding biomolecular (FKBP) or any binding pair of             biomolecular interaction domains are collectively a             cyclophilin-Fas fusion biomolecular (CyP-Fas) and a FK506             binding biomolecular (FKBP);         -   g. wherein the biomolecular interaction domain(s) is a             calcineurin A (CNA) and/or a FK506 binding biomolecular             (FKBP) or any binding pair of biomolecular interaction             domains are collectively a calcineurin A (CNA) and a FK506             binding biomolecular (FKBP);         -   h. wherein the biomolecular interaction domain(s) is a             gibberellin insensitive (GIA) and/or a gibberellin             insensitive dwarf1 (GID1) or any binding pair of             biomolecular interaction domains are collectively a             gibberellin insensitive (GIA) and a gibberellin insensitive             dwarf1 (GID1);         -   i. wherein the biomolecular interaction domain(s) is a             Snap-tag and/or a Halo tag, or any binding pair of             biomolecular interaction domains are collectively a Snap-tag             and a Halo tag;         -   j. wherein the biomolecular interaction domain(s) is a             T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52),             or any binding pair of biomolecular interaction domains are             collectively a T14-3-3-cdeltaC and a C-Terminal peptides of             PMA2 (CT52);         -   k. wherein the biomolecular interaction domain(s) is a PYL             and/or a ABI, or any binding pair of biomolecular             interaction domains are collectively a PYL and a ABI;         -   l. wherein the biomolecular interaction domain(s) is a             nucleotide tag and/or a zinc finger domain, or any binding             pair of biomolecular interaction domains are collectively a             nucleotide tag and a zinc finger domain;         -   m. wherein the biomolecular interaction domain(s) is a             nucleotide tag, or any binding pair of biomolecular             interaction domains are collectively a pair of nucleotide             tags;         -   n. wherein the biomolecular interaction domain(s) is a             Fluorescein isothiocyanate (FITC) and/or a FITC binding             biomolecular or any binding pair of protein interaction             domains are collectively a FITC and a FITC binding protein;             and/or         -   o. wherein the protein interaction domain(s) is a             (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein             or any binding pair of protein interaction domains are             collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE             binding protein.     -   10. The composition of paragraph 9, wherein the nucleotide tag         is a DNA tag or dsDNA tag.     -   11. The composition of paragraph 2, further comprising a cell         expressing or comprising the signaling polypeptide.     -   12. The composition of paragraph 11, wherein the TCR recognition         domain is allogeneic, autologous, or xenogenic to the cell.     -   13. The composition of paragraph 11, wherein the TCR recognition         domain is synthetic.     -   14. The composition of paragraph 11, wherein the TCR recognition         domain comprises a MHC and a peptide, wherein the peptide is         allogeneic, autologous, or xenogenic to the cell.     -   15. The composition of paragraph 11, wherein the TCR recognition         domain comprises a MHC and a peptide, wherein the peptide is         synthetic.     -   16. The composition of paragraph 11, wherein the cell is a NK         cell, dendritic cell, regulatory T cell, or effector T cell.     -   17. The composition of paragraph 11, wherein the cell is further         engineered to knockdown the native MHCI/II expressed on the cell         surface.     -   18. A method of treating or preventing an autoimmune disease or         condition; T cell mediated inflammation or immune response;         malignant T cell condition; transplant rejection; or GvHD in a         subject in need thereof, the method comprising administering to         the subject a composition of paragraph 1.     -   19. The method of paragraph 18, wherein the transplant is an         allogeneic hematopoietic stem cell or solid organ         transplantation.     -   20. The method of paragraph 18, wherein the malignant T cell         condition is T cell acute lymphoblastic leukemia or T cell         lymphoblastic lymphoma.     -   21. The method of paragraph 18, wherein the autoimmune disease         is type 1 diabetes, vitiligo, multiple sclerosis, alopecia,         celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.     -   22. The method of paragraph 18, wherein the T cell mediated         immune response is an anti-drug specific response to a biologic,         cell therapy, and/or gene therapy.     -   23. The method of paragraph 22, wherein the biologic,         cell-therapy, or gene therapy is an adeno-associated virus (AAV)         gene therapy, a genome editing agent, or enzyme replacement         therapy.     -   24. The method of paragraph 18, wherein the disease is type 1         diabetes and the TCR recognition domain comprises sequences with         at least 95% sequence identity to: one or more of SEQ ID NOs:         8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and         2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs:         20128-2129.     -   25. The method of paragraph 18, wherein the disease is vitiligo         and the TCR recognition domain comprises sequences with at least         95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22;         or HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO:         2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101         and SEQ ID NO: 2021.     -   26. The method of paragraph 18, wherein the method is a method         of treating and/or preventing GvHD and the TCR recognition         domain comprises sequences with at least 95% sequence identity         to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or         HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO:         2039.

EXAMPLES Example 1

Auto- and allo-reactive T cells attacking patient's or donor's cells or organs is the major cause of autoimmunity and transplant rejection. Current treatments involve stringent immunosuppressant therapy, which can lead to severe side effects. Specifically, depletion of auto and alloreactive T cells prevents autoimmunity and transplant rejection without immunosuppressant modalities. T cells engineered with a Chimeric antigen ligand (CAL) can redirect their specificity toward the pathologic T cells. T cells engineered with a Chimeric antigen receptor (CAR) can redirect their specificity and have already been approved to treat some types of B cell malignancies. Currently, engineered regulatory T cell, which can inhibit immune reactions in an antigen-dependent manner, is under investigation to expand the application of CAR T cells therapies such as autoimmune disease and transplant rejection.

To specifically deplete disease causing T cells, described herein is a CAL and/or CAR that can recognize and bind to the specific T cell receptor on the disease causing T cells. This CAL and/or CAR is a composed of a peptide-HLA (e.g., a monomer or multimer or oligomer thereof) as the recognition domain fused to signaling domains from T cell receptors. A split version of the system can also be generated where the system is composed of two pieces. One piece is an universal CAR with T cell signaling domains as the intracellular portion and a biomolecular interaction domain as the extracellular domain. The second piece is an adaptor molecule, e.g, a CAL composed of a peptide-HLA oligomers (or monomer or multimer) fused to the cognate biomolecular interaction domain.

Most CAR T cells therapies for autoimmune disease and transplant rejection uses regulatory T cells and are designed to target the organ to provide organ-specific, e.g., local immunosuppression. In contrast, the present, e.g., CAL, design is targeting the source immune cells inside that patient that are causing the disease. Accordingly, provided herein is a cell-based therapy for autoimmune diseases and transplant rejection.

Example 2

Dendritic cells (DCs) are the interconnection between innate and adaptive immune system and are able to induce protective immune responses following stimulation by a variety of stimuli. Based on their phenotype and function, DCs can be divided into conventional DCs and plasmacytoid DCs (pDCs). Many studies have shown that DCs are essential mediators of pro-inflammatory or anti-inflammatory (tolerogenic) responses. The subsets of DCs that suppress immune responses are generally known as tolerogenic DCs because of their functions in inducing T cell apoptosis, anergy and regulatory T cells (Tregs).

A tolerogenic state in DCs (tol-DC) can be induced using several pharmacological agents, such as cyclosporine A, rapamycin, dexamethasone, vitamin A, vitamin D or other cytokines and growth factors. Recently, the insertion of exogenous DNA to enhance tol-DC function has been investigated as a therapeutic possibility for treating autoimmune diseases. ‘Killer’ DCs, obtained by transfection with DNA encoding FasL or TNF-related apoptosis-inducing ligand, efficiently induce T-cell apoptosis and prevent the rejection of heart grafts in animal models.

However, the effect of these DCs are not specific. Overexpression of inhibitory molecules, including IL-10, TGF-β, CTLA-4 and SOCS1, enable tol-DCs to more efficiently induce Tregs which might cause systemic immune suppression.

Alloreactive immune cells can be suppressed in a specific manner using the methods described herein, e.g., by using a universal UniCAR DC system that presents the donors' pMHC tetra/dextramers (e.g., the CAL) and binds to a genetically engineered DC (K/O MHC I/II but presents donor peptide on the tetra/dextramer). In some embodiments, the pMHC can be provided as a monomer, oligomer, or multimer.

Any form of the recipient pMHC (e.g., monomer/oligomer/tetramer/dextramer)+ targeting module (e.g, the CAL) can be used with CAR-DC for the suppression of allo-reactive T-cells in the recipients. A library of identified different alloantigens+correlated or associated with MHC monomers/oligomers/tetramers/dextramers (e.g. HLA-A2+insulin peptide) and attached to the targeting molecules of the CAL and/or CAR system can be produced. Such a library can be commercially available to be combined with DCs for targeting and destroying the auto reactive T-cells.

Cells can be produced that are genetically modified with the specific gene deletion of some genes (IL-12 and NF-κB, MHC I and MHC II [The targeting module will provide these cells with the donor's MHCs]) and insertion of some other genes (IL-10, TGF-β, CTLA-4 and SOCS1). These cells can be commercially ready to use and could be combined with the specific donor, pMHC and targeting module chosen from the aforementioned library.

Example 3

Alloreactive memory T cells have been shown to have the key role in the activation of the recipient immune system and rejection of the allograft. They are known to be the main barrier for tolerance induction through mixed chimerism and other strategies. These cells are very resilient and not responsive to the preconditioning protocols and they recover from irradiation and T cell depletion rapidly. By using CAR-T-effector (Conventional, Supra, Universal CAL or CAR) and attaching any form of the donors' pMHC (monomer, oligomer, tetramer or dextramer) (e.g., a CAL) to the system, the resulting compositions of the invention can be used to decrease or eliminate the need for improved immunosuppressant therapy in the context of transplantation. The engraftment of hematopoietic stem cells can achieve durable mixed chimerism with minimal or no need for toxic preconditioning protocols.

If donor pMHC (monomer/oligomer/tetramer/dextramer; e.g., a CAL) is used in combination with CAR T effector for the abrogation of the allo/xeno reactive T-cells in the recipient, the need for immunosuppressants would be decreased or eliminated. A library of different Donor MHC monomers/oligomers/tetramers attached to the targeting molecules (e.g., the CALs) of the SUPRA/UNI/universal CAL and/or CAR system can be produced to be commercially available. A shelf ready library of the most prevalent MHC (+immunologic wildtype or synthetic peptide molecules that could be used to be combined with the MHC) monomers/oligomers/tetramers could be commercially produced and offered to be used with this system.

Example 4

The involvement of the adaptive immune system in auto-immune diseases has been extensively characterized. T cells are critical contributors to autoimmune diseases. Conventional T (Teff or T helper) cell subsets that play a role in B cell activation and differentiation produce various inflammatory cytokines and destroy target cells with direct cytotoxicity. CAR-T cells have been used to destroy autoimmune B and T cells in a fashion similar to the way in which CD19CAR T cells target and destroy leukemia cells. Targeting autoreactive memory T and B-cells has shown some results. As these methods are nonspecific, they induce some extent of generalized immune suppression and they are not completely effective. The correlation of particular peptide+MHC molecules with certain autoimmune conditions have been expansively studied. By using the available human/animal model pMHC monomer/oligomer/tetra/dextramers and combining them with the SUPRA/UNI/universal CAL and/or CAR technology, we are able to target the autoreactive immune cells in a highly specific manner.

If any form of auto-antigen on pMHC (monomer/oligomer/tetramer/dextramer) (e.g., the CAL) is used in combination with CAR T effector/Treg to abrogate autoreactive T-cells in the recipient, the need for immunomodulatory drugs would be decreased or eliminated. A library of identified autoantigens and correlated MHC monomer/oligomer/tetra/dextramers (e.g. HLA-A2+insulin peptide) attached to the targeting molecules of the SUPRA/UNI/universal CAL and/or CAR system can be produced to be commercially available to be combined with ideal SUPRA or universal CAR T-reg/T effector for targeting and destroying the autoreactive T-cells.

Example 5

Although new advances have increased survival after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) is still the leading cause of late morbidity and mortality after transplant. Current treatment choices are limited in efficacy specifically in steroid-refractory disease, and there is no robust data to help with management decisions.

Adoptive T cell therapy (ACT) refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CAR) constitute the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemias and lymphomas and have produced remarkable results in the clinic. The technology described herein permits the opportunity to target diseased cells with specific antigens or receptors very accurately. In the context of GvHD, universal UNICAR T cells can be designed to find the recipient reactive T cell in the donor T cell populations. If the pMHC of the recipients is fused to a Target Module that binds to the UniCAR (e.g, to form a CAL), this system can recognize the TCR repertoires in the donor T/B cell population that can bind to those MHCs. If the recipients' pMHCs are fused to a target module that binds to the universal CAR, this system can recognize and bind to the T cell repertoires in the donor T cell population that can bind to those MHCs.

If the recipient pMHC (monomer/oligomer/tetramer/dextramer) (e.g., a CAL) is used in combination with CAR T effector/Treg for the abrogation of the recipient reactive T-cells in the donor HSC, these reactive T-cells will be depleted and GvHD would not happen. A library of identified different antigens+correlated human/animal models MHC tetramers/dextramers (e.g. HLA-A2+ peptide) attached to the targeting molecules of the SUPRA/UNI CAR system can be produced to be commercially being available to be combined with ideal SUPRA CAR T-reg/T effector for targeting and destroying the reactive T-cells against the recipients. A library of wild type and/or synthetic MHC monomers/oligomers (e.g. HLA-A2+ peptide) attached to the targeting molecules of the universal CAL system can be generated to be mixed with universal CAL T-reg/T effector for targeting and killing the reactive T-cells against the recipients within the donor T cell population.

Example 6

Although new advances have increased survival after allogeneic hematopoietic stem cell transplantation (HCT), chronic graft-versus-host disease (GvHD) is still the leading cause of late morbidity and mortality after transplant. Current treatment choices are limited in efficacy specifically in steroid-refractory disease, and there is no robust data to help with management decisions.

Adoptive T cell therapy (ACT) refers to the therapeutic use of T cells. T cells genetically engineered to express chimeric antigen receptors (CAR) constitute the most clinically advanced form of ACT approved to date for the treatment of CD19-positive leukemias and lymphomas and produced remarkable results in clinical. This technology provides the opportunity to target the cells with specific antigens or receptors very accurately. In the context of GvHD, UNICAR T cells can be designed to find the recipient reactive T cell in the donor T cell populations. If the pMHC of the recipients be fused to a Target Module that bind to the UniCAR, this system can recognize the TCR repertoires in the donor TB cell population that can bind to those MHCs.

If the recipient pMHC (tetramer/dextramer) is used in combination with CAR T effector/Treg for the abrogation of the recipient reactive T-cells in the donor HSC, these cells will be depleted and GvHD would not happen. A library of identified different antigens+correlated human/animal models MHC tetramers/dextramers (e.g. HLA-A2+ peptide) attached to the targeting molecules of the SUPRA/UNI CAR system can be produced to be commercially being available to be combined with ideal SUPRA CAR T-reg/T effector for targeting and destroying the reactive T-cells against the recipients.

Example 7—Engineered Lymphocytes for Prevention of Pediatrics Vascularized Composite Allograft Rejection Engineered

Described herein is the development of a clinically applicable tolerance-inducing regimen for VCA transplantation through the establishment of stable mixed chimerism, augmented by the state-of-the-art CAL and/or CAR T cell adoptive immunotherapy and synthetic biology.

Mixed chimerism in animal models and human allograft recipients have only shown to be transient, suggesting that tolerance here relies on the peripheral inactivation of donor-specific T cell. The difficulty in achieving durable mixed chimerism and long-term graft acceptance is the presence of high levels of alloreactive memory T cells that are known to hinder tolerance induction in sensitized rodents, NHPs, and human. The present approach to achieving stable mixed chimerism is to utilize advanced engineering techniques to generate CAL and/or CAR T cell therapeutics to specifically delete the recipient's alloreactive memory immune cells that are reactive against the donor bone marrow and tissue cells to help with engraftment of HSCT and stable mixed chimerism. Selective depletion of memory T lymphocytes with CAL and/or CAR-T cell therapy can help to achieve durable mixed chimerism and tolerance which eventually leads to an immunosuppressant free regimen. Currently this challenge is addressed by administering several immunosuppressant combinations in combination with intense whole-body irradiation which result in depletion of all the immune cells as a consequence of nonspecific targeting. This shotgun approach imposes a severe immune compromised state to the recipients which subsequently brings myriads of consequences including opportunistic infections and malignancies. The technology herein can delete specifically memory T cells (Aim 1) or only alloreactive memory T cells (Aim 2):

Aim 1: It has been shown that CD127+/CD45RO+ memory T cells have a significant role as central, effector and stem cell memory T cells and are known as the most potent constituents of the alloreactive T cell repertoire. These cells have been shown to be the major contributor in chronic rejection of the allograft. To address the challenge of memory T cells a CAL and/or CAR T-effector cell can be generated with scfv against two general markers of memory T cells (CD127+ and CD45RO+). This will be followed by the induction of mixed chimerism protocol in a double humanized mouse model. The results from this aim will show depletion of memory T-cells increases the efficacy of mixed chimerism protocol by improving the engraftment of hematopoietic stem cells hence helping to eliminate or decrease the need for immunosuppressant in mixed chimerism induction protocols.

Aim 2: Due to the diversity of antigen-specific T cells in the context of transplantation, a CAL and/or CAR system is provided that has the flexibility to locate and attack different alloreactive T cells simultaneously. CAL and/or CAR T-eff cells with donor pMHC can target alloreactive T-cells that have TCR against the donor MHCs. We will start with single and double specific antigen-MHC systems. Then, we will utilize a peptide library that is generated from the donor's allograft peptidome to be loaded on commercially-available, exchangeable peptide-MHC multimers. The combination of these pMHCs and CAL and/or CAR T cells will be used to target and destroy alloreactive T cells of the recipients. This development provides a flexible CAL and/or CAR design that can target antigen specific alloreactive T cells.

Described herein is the investigation of a novel adaptive immune cell therapy strategy, making use of the CAL and/or CAR T-cell technology, for VCA tolerance induction.

Example 8—Engineered Lymphocytes for Prevention of Pediatrics Vascularized Composite Allograft Rejection

Significance: There are nearly 2 million people living with limb loss in the US, where over 185,000 amputations occur each year, most of which are sustained by victims of burn injuries, traffic accidents, and medical conditions¹. 112600 children with amputations were treated in US emergency departments alone from 1990 to 2002. Children are frequently the victims of severe burns with limb loss² and they will suffer from its consequences for the rest of their lives. Over the last decade, vascularized composite allotransplantation (VCA), the transplantation of limbs and face from a deceased donor, has become a good alternative for the reconstruction of devastating injuries of these specialized tissues. VCA represents a unique new treatment option for severe soft tissue defects following burn injury to achieve both psychosocial and functional rehabilitation³. The world's first pediatric bilateral hand transplant was successfully performed between an unrelated donor-recipient pair in 2016⁴. The shortcoming is that despite the use of potent immunosuppressive drugs, acute rejection of “foreign” VCA occurs in up to 90% of patients⁵. Increased doses of immunosuppression, with numerous life-threatening complications, is the current approach to prevent loss of the VCA⁵.

During the course of evolution, our immune system has gained the ability to recognize self from nonself cells and attack the “foreigners” by cells such as T Lymphocytes. The Human Leukocyte Antigen (HLA) system (Major Histocompatibility Complex [MHC] in human) comprises cell surface molecules specialized to present antigenic peptides to the T-cell⁶⁷. These peptides are known as Minor Histocompatibility Antigens (MiHA). MiHA sequences can differ among individuals and many of these differences can be recognized by T cells, hence causing the initiation of the rejection process. T-cells are trained to recognize self from non-self MHC/peptides. Each person can present a combination of 12 different MHC class I/II on the surface of his/her cells. (HLA) typing is used to match recipients and donors MHCs for transplants. A close match between a donor's and a recipient's HLA markers is essential for a successful transplant outcome. A potential donor must match a minimum of 6-8 HLA markers which makes finding an ideal donor very difficult. Even after finding the semi ideal donor these patients will be on immunosuppressants for the rest of their life⁸. While these drugs are generally effective, the sequelae of such chronic immunosuppression are well known, and most recipients continue to develop myriad side effects and complications, including opportunistic infections, multiple organ dysfunction, and malignancies.

Transplantation tolerance that allows for the elimination of immunosuppressive drugs, has been the “holy grail” for transplantation medicine since its beginnings⁹. Developing a reproducible, safe tolerance induction protocol would expand VCA use in burn patients as well as in congenital anomalies as demonstrated by the successful transplantation of the arm and hand⁶ and the lower extremity¹⁰. The extension into other organ transplants is also a clear direction to broaden the significance. The overall goal of this research is to enable the clinically applicable tolerance-inducing regimen for VCA transplantation through the establishment of stable mixed chimerism, augmented by the state-of-the-art adoptive immunotherapy and synthetic biology.

Tolerance can be induced through the development of a mixed chimerism protocol for VCA transplants in partially mismatched subjects^(8,11). Mixed chimerism is the use of a donor hematopoietic stem cell transplant (HSCT) along with a VCA to induce a hybrid immune system in the recipient that recognizes the donor VCA as “self” and therefore does not reject it. Mixed chimerism in nonhuman primates (NHPs) and human allograft recipients has only shown to be transient, suggesting that tolerance here relies on the peripheral inactivation of donor-specific T cell. The difficulty in achieving durable mixed chimerism and long-term graft acceptance is the presence of high levels of alloreactive memory T cells that are known to hinder tolerance induction in both sensitized rodents^(12,13) and NHPs^(14,15). The present approach to achieving stable mixed chimerism is to utilize advanced engineering techniques to generate T cell therapeutics to specifically delete the recipient's alloreactive memory immune cells that are reactive against the donor bone marrow and tissue cells.

Cellular Immunotherapy

Recently, T cells that are genetically engineered to express chimeric antigen receptor (CAR) produced remarkable results for the treatment of CD19-positive leukemias and lymphomas, leading to complete remissions in pediatric patients^(16,17) and evidencing that immune cells can be targeted and eliminated by CAR T cell technology. Thus far, CAR T therapeutics have led to two FDA-approved therapies—the two medication were licensed by Novartis and Gilead in the value 9 and 12 billion dollars¹⁸. First-generation CARs contained only an extracellular antigen-binding domain, a transmembrane domain, and the signaling domain of CD3z. In later generation CARs, intracellular costimulatory domains, derived from either CD28/4-1BB were added to enhance proliferation, persistence, and activity¹⁹. Recently, to expand the capability of CAR T cells, we introduced a split, universal, and “programmable” (SUPRA) CAR system that simultaneously encompasses multiple critical features²⁰. This system has the ability to switch targets without reengineering the T cells, finely tune T cell activation strength, and sense and respond to multiple antigens. These features make the split CAR-T technology uniquely suitable for targeting alloreactive T-cells in recipients and enhancing transplantation tolerance.

Mixed Chimerism and Tolerance Induction in VCA

Large animal models of Mixed Chimerism: Our laboratory achieved transient mixed chimerism in two-haplotype full-mismatch MGH miniature swine after musculocutaneous VCA and nonmyeloablative recipient conditioning (total body irradiation (TBI) and transient T-cell depletion) concomitant to bone marrow transplantation (BMT)²¹. While the immunomodulatory effect of BMT was demonstrated through in vitro unresponsiveness to the donor, the chimerism levels fell rapidly following cessation of immunosuppression with resulting VCA rejection and loss. This was the first experimental VCA study to demonstrate that transient chimerism alone is neither sufficient for achieving transplant tolerance nor prolonging VCA survival.

Recently, in order to progress towards clinical translation, the previous Mixed Chimerism Induction Protocols (MCIP) in miniature swine were modified to remove T-cell depletion completely in exchange for a higher dose, TBI and TI commencing two days prior to surgery. Following osteomyocutaneous VCA, a pen-operative course of co-stimulatory blockade was given (on POD0, 2, 4 and 6) in addition to a 30-day regimen of tacrolimus (target levels: 10-15 ng/mL) before gradual taper to discontinuation on POD 45²². With this MCIP, we were indeed able to achieve stable mixed chimerism (n=2) following donor BMT without the development of split tolerance in MHC class I mismatched/class II matched recipients and have successfully withdrawn all immunosuppression for >100 days. Building on these results, the next iteration towards clinical translation of these studies is to decrease the toxicity of current protocol to be able to extend it for achieving stable mixed chimerism across full MHC-mismatch barriers—which can utilize immunomodulatory strategies such as CAL and/or CAR T cell therapy for depletion of alloreactive memory T cells that may eventually result in elimination or decrease in the immunosuppressive regimen for VCA transplantation.

Small animal models of Mixed Chimerism: The principles that underlie tolerance induction through mixed chimerism induction protocol are the same for all species. Briefly, mice can be conditioned with intraperitoneal mAb injections including, anti-CD8 and anti-CD40L, anti-CD154, costim. blockade (on day 0 with respect to the BMT). This will be combined with, TBI to be given 6 h before injection of 2.5 10⁵ bone marrow cells (BMC). The compositions and methods described herein can be used for improving mixed chimerism induction protocol in mice models.

Described herein is a CAL and/or CAR designed to target and destroy alloreactive memory T cells thereby improving the engraftment of hematopoietic stem cells and achieving durable mixed chimerism with minimal or no need for toxic preconditioning protocols. To test this multi-pronged approach, we will first evaluate two different strategies of targeting alloreactive memory T cells. The first approach is to design a CAL and/or CAR system that will specifically attack memory T cells through surface markers that uniquely identify this cell population regardless of their TCR specificity. This approach will spare the other immune cells, and, thus retain a healthy immune system.

A second approach is to design CAL and/or CARs that specifically bind to alloreactive TCRs on the recipient's T cells. To do so, instead of using a single-chain variable fragments (scFvs) that is in all classical CAR T cells, we will use peptide-MHC multimers as the ligand recognition domain on the CAL and/or CAR. However, due to a large number of unique alloreactive TCR, many different CAL and/or CAR/pMHCs are needed. This poses a challenge in designing and introducing these CAL and/or CARs into T cells. To circumvent this challenge, we propose to use a two-component split CAL and/or CAR design composed of universal CAL and/or CAR and an adaptor molecule that bridges the CAL and/or CAR T cells to alloreactive T cells. The universal CAL and/or CAR comprises a scFv targeting FITC fused to TCR signaling domains. The adaptor molecules consist of pMHC tetramer fused to a FITC. The addition of pMHC tetramer will bind to the alloreactive TCR and recruit the universal CAL and/or CAR T cell to kill alloreactive T cells. This design allows us to be able to target many variants of TCR with only one CAL and/or CAR. Next, the donor MHCs will be loaded with donor tissue specific peptides that will be generated by utilizing previously published protocols. We can first design CAL and/or CARs to target mouse MHC in a humanized/skin graft mouse model.

Data-Targeting Alloreactive T-Cells with Peptide-MHC Tetramers

The following have been addressed by in vitro data: 1. Whether targeting a memory T cell by using CAR T+pMHC multimer is obtainable. In this embodiment, the anti-FITC CAR expressed by the Jurkat cells is a CAR and the pMHC multimer is a CAL.

-   -   A. Would the Tetramer efficiently bind to the CAL and/or CAR and         target T cells?     -   B. Would the Jurkat CAL and/or CAR T cells get activated by         attaching to the adaptor+ Target     -   C. Would the Target OT I cells get activated and try to kill the         Killer Target T cells?         2. Whether killing a memory T cell with primary CD8 T cell+pMHC         multimer would be feasible?

1. Peptide-MHCs Tetramers Activate Jurkat Cells but not the Target Cells

We used anti-FITC CAR Jurkat cells. These cells are an immortalized line of T lymphocyte cells that are used to study activation, signaling, and expression of various chemokine receptors of T cells. Jurkat cells cannot act as a killer cell but will respond to and get activated by the ligands in the same way as actual T cells. FIG. 1 shows tetramers attaching to and staining OTi target T cells in a dose dependent fashion.

The anti-FTC CAR Jurkat cells used in these experiments comprise the following nucleotide sequence and express the following polypeptide sequence.

DNA sequence of anti-FITC CAR. Nucleotides 1-9 =  Kozak sequence. Nucleotides 10-78 = CD8a leading  peptide. Nucleotides 79-831 = anti-FITC ScFv.  Nucleotides 832-861 = myc. Nucleotides 874- 1008 = CD8  hinge. Nucleotides 1009-1211 = CD28. Nucleotides 1212-1338 = 4-1BB. Nucleotides 1339- 1677 =CD3ζ. Nucleotides 1678-2388 mCHERRY. SEQ ID NO: 2747 gccgccaccATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTT GCTGCTCCACGCCGCCAGGCCGGCTGCAcaagttcagctcgttgaatccg gcggaaaccttgttcaaccggggggttccctccgccttagttgtgccgca tctggttttacgtttggatcattttccatgtcatgggtgcggcaggctcc cggggggggactcgaatgggttgcgggtcttagcgcccgatcaagcctca ctcactatgcagatagcgtaaaaggcaggtttacaatctcacgagacaac gcgaagaactcagtctatcttcagatgaactctctccgagtcgaggatac ggctgtctattattgtgcacgccgctcctacgattcctcaggctacgccg ggcacttttattcatacatggatgtctggggccagggaaccttggtaact gtgtctggaggaggtggatcagggggtggcggaagtggcggaggtggatc ctccgtcttgacccagccctcctcagtcagtgctgcccctggccaaaagg ttacaatatcatgttcaggtagtacgtcaaacataggcaacaactacgtg agttggtaccagcaacatcctggcaaagcacctaagttgatgatctatga tgtcagtaagcggccatctggggtacccgaccgattctcaggctcaaaga gcggaaactccgcgtccctcgacataagtggcctccaatccgaggacgag gccgactactactgtgctgcttgggacgactctctctcagagtttttgtt tgggaccggtacaaaactgactgttcttgggGAACAAAAACTCATCTCAG AAGAAGATCTGAATGGGGCCGCAACCACGACGCCAGCGCCGCGACCACCA ACACCGGCGCCCACCATCGCGTTGCAGCCCCTGTCCCTGCGCCCAGAGGC GTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCG CCTGTGATTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGCTTGCTAT AGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAG GAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCG GGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCA GCCTATCGCTCCAAACGGGGCAGAAAGAAACTCCTGTATATATTCAAACA ACCATTTATGAGACCAGTACAAACTACTCAAGAGGAAGATGGCTGTAGCT GCCGATTTCCAGAAGAAGAAGAAGGAGGATGTGAACTGAGAGTGAAGTTC AGCAGGAGCGCAGACGCCCCCGCGTACAAGCAGGGCCAGAACCAGCTCTA TAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGA GACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCT CAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTA CAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATG GCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTT CACATGCAGGCCCTGCCCCCTCGCGGCatgGTGAGCAAGGGCGAGGAGGA TAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGG GCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGC CCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCC CCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCA AGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCC TTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGG CGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCT ACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATG CAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGA GGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACG GCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCC GTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTC CCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCC GCCACTCCACCGGCGGCATGGACGAGCTGTACAAGtaa Peptide sequence of anti-FITC CAR. Residues  1-21 = CD8a leading peptide. Residues 22-274 = anti-FITC ScFv. Residues 275-284 = myc. Residues  289-333 = CD8 hinge. Residues 334-401 = CD28. Residues 402-443 = 4-1BB. Residues 444-556 = CD3ζ. Residues 557-792 mCHERRY. SEQ ID NO: 2748 MALPVTALLLPLALLLHAARPAAQVQLVESGGNLVQPGGSLRLSCAASGF TFGSFSMSWVRQAPGGGLEWVAGLSARSSLTHYADSVKGRFTISRDNAKN SVYLQMNSLRVEDTAVYYCARRSYDSSGYAGHFYSYMDVWGQGTLVTVSG GGGSGGGGSGGGGSSVLTQPSSVSAAPGQKVTISCSGSTSNIGNNYVSWY QQHPGKAPKLMIYDVSKRPSGVPDRFSGSKSGNSASLDISGLQSEDEADY YCAAWDDSLSEFLFGTGTKLTVLGEQKLISEEDLNGAATTTPAPRPPTPA PTIALQPLSLRPEACRPAAGGAVHTRGLDFACDFWVLVVVGGVLACYSLL VTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYR SKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRS ADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEG LYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQ ALPPRGMVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYE GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPE GFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKK TMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQL PGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK

Next, we showed that binding of adaptors to the OTi target T cells would not activate them. To evaluate that, we looked at CD69 expression which is a general activation marker for T cells. Interestingly, although adaptor binds to and stains target T cells (FIG. 1) it did not cause activation of OTi target T cells (FIG. 2).

B. Subsequently, we studied the activation of Jurkat CAR T cells after binding to the adaptors. As shown in FIG. 3, binding of adaptor to the Jurkat T cells causes high expression of CD69 at all measured time points.

This activating binding of Jurkat cells-pMHC is in contrary to the binding of OTi (target)-adaptor (FIG. 2). The best explanation for this interesting finding may lie behind the fact that FITC/anti-FITC binding between Jurkat-pMHC is much stronger than OT I-pMHC as the first one is a high affinity antigen-antibody binding and the second one is a weaker TCR-pMHC binding. This difference in the strength of binding between CAR-adaptor and Target-adaptor makes this novel approach an ideal setting for the purpose of targeting memory T cells.

C. We also looked at the Jurkat cells count after 24 hrs. as an indicator of their condition and whether this activation would cause any cell death indicated by a decrease in their numbers (FIG. 2).

2. Primary CD8 Anti-FITC CAR T Cells can Specifically Target and Kill Alloreactive OTi Cells.

The previous set of experiments with Jurkat+adaptor+OTi target cells confirmed the feasibility of targeting alloreactive T cells by adaptor molecules and showed activation of CAR T cells against the target cells. Next, we wanted to study whether primary CD8 CAR T killer cells+adaptor molecules can be used not only to identify but also to kill the OTi T cells. As shown in the previous set of experiments the binding between pMHC and target cell was not causing activation of target T cells in contrary to the adaptor (FITC+pMHC, e.g., the CAL)-CAR T cells binding Hence, we used human primary CD8 T cells to see whether the strength of CAR T+adaptor (e.g, CAL)+target is enough to initiate and accomplish the killing process. Different concentrations of adaptor+CD8 CAR T were co-cultured with OTi T cell to see the activation and cytotoxicity of each T cell type. As shown in FIGS. 4 and 5, CD8 CAR T cells cytotoxicity on OTi cells is dependent on the adaptor (e.g., CAL) concentration and was over to 50% with the maximum dose that we used. Additionally, this cytotoxicity is highly specific. (FIG. 5) Human CD8 anti-FITC CAR T cells were used here. Similar validations can be conducted with mouse anti-FITC CARs in both in vivo and in vitro experiments.

Described herein is a novel method to use pMHC multimers for specifically targeting and destroying allo/autoreactive T cells responsible for graft rejection and autoimmune diseases. Our approach is the first demonstration of utilizing pMHC multimer (e.g, CAL) in combination with CAR T cell technology for specific targeting of alloreactive T cell population. We believe this approach is the most specific strategy for eliminating the destructive role of alloreactive T cells in transplantation. The potential application of this method extends far beyond elimination of immunosuppressants in transplantation rejection and GVHD treatment and could be applicable to many autoimmune conditions including type 1 diabetes, multiple sclerosis, rheumatoid arthritis, scleroderma, and myriad disorders that originate from auto reactive T cells dysfunction.

Provided herein is a novel approach for targeting the memory repertoire with anti CD45RO/CD127 with CAL and/or CAR T cell therapy. Although this strategy might not be as specific as using pMHC multimers, it still conveys a significant novelty. By depleting the memory T cells population there is much less of a need for induction protocols.

Also contemplated herein are individualized peptide libraries based on the differences between the recipient and the donor proteome and immune-peptidome. Hundreds to thousands of MHC-associated peptides can now be identified in a single measurement using optimal biological model systems. As mentioned earlier, studies involving MHC identical grafts indicate that minor histocompatibility antigens may also mediate rejection. Human Immuno-Peptidome Project (HIPP) is an international project that was created to accelerate research toward robust and comprehensive analysis of immunopeptidome. HIPP has published the technical guidelines that represent the information required to sufficiently conduct and interpret all of the immunopeptidomics experiments. These nonhomogeneous peptides can potentially initiate rejection through activating alloreactive T cells.

By utilizing these peptides in combination with exchangeable MHC tetramer systems, we can employ these peptides to identify and target donor reactive T cells. The default peptide on exchangeable peptide MHC systems can be removed by UV light exposure and replaced by any desired peptide. The donor multimers MI-ICs can be generated using commercially available services, unloaded with UV and then loaded with donor tissue specific peptides that will be generated by utilizing previously published protocols. By utilizing these peptides in combination with exchangeable MHC tetramer systems, we can employ these peptides to identify and target donor reactive T cells.

As described herein, selective depletion of memory T cells with CAR-T cell therapy can help in achieving durable mixed chimerism and tolerance which lead to the elimination or decrease in immunosuppressive regimens. Currently this challenge is addressed by administering intense whole-body irradiation with several immunosuppressant combinations for targeting all the immune cell populations. This shotgun approach results in severe immune compromised state in the recipient which subsequently brings myriads of side effects and complications, including opportunistic infections, reno-vascular dysfunction, and malignancies.

Memory T cells (CD127+/CD45RO+) are known to be the main contributor in rejecting allografts 2425. Hence, their depletion is expected to help engraftment of HSC and increase the survival of allografts. To investigate this hypothesis, we will generate CAL and/or CAR T-effector cell with scfv against a general marker of memory T cells (CD127+ and CD45RO+). This will be followed by induction of mixed chimerism protocol in a humanized mice model. The engraftment and stability of mixed chimerism will be followed by the presence of donor lymphoid and myeloid cells in the recipient blood stream until the study endpoints.

Activity 1.1 Generation and evaluation of AND logic CAL and/or CAR T cells to only target cells co-expressing both CD127+ and CD45RO+. Since no single surface marker can uniquely specify memory T cells, we will need to target a combination of CD markers to ensure specificity. We chose CD127 and CD45RO because they have proven to uniquely identify memory T cells 2627. To accomplish this combinatorial targeting, we will leverage an advance CAR system called SUPRA. This (SUPRA) CAR is a two-component receptor system comprises a universal receptor (zipCAR) expressed on T cells and a targeting scFv adaptor (zipFv). The zipCAR universal receptor is generated from the fusion of intracellular signaling domains (FIG. 6) and a leucine zipper as the extracellular domain. The zipFv adaptor molecule is generated from the fusion of a cognate leucine zipper and an scFv. The scFv on the zipFv binds to the antigen, while the leucine zipper binds to and activates the zipCAL and/or zipCAR on the T cells. For transplantation immunotherapy, these features can mitigate over-activation and enhance specificity.

The SUPRA CAL and/or CAR system can also be designed to perform AND combinatorial logic of antigen recognition. Orthogonal leucine zipper pairs can be used to generate CAL and/or CARs with split signaling domains (e.g., CD3z, CD28, 4-1BB), thus enabling independent and simultaneous control of these pathways (FIG. 6B). T cells need both CD3z and costimulatory signaling simultaneously to be fully activated therefore, each CAL and/or CAR can be readily paired with scFvs that target different antigens, thereby enabling two antigen combinatorial and logical antigen sensing.

Using the SUPRA system in human primary T cells, we have demonstrated that the activity of SUPRA CARs can be finely regulated via multiple mechanisms to limit over-activation (FIG. 7). SUPRA CAR T cells can also logically respond to multiple antigens for improved target specificity. Furthermore, orthogonal SUPRA CARs can be used to inducibly regulate multiple signaling pathways or T cell subtypes to increase the breadth of immune responses that can be achieved. A summary of the SUPRA CAL and/or CAR T cell features used for this project can be found in FIGS. 6A-6B, 7A-7D.

zipCAR Receptor Construct Design: As described herein, AND logic CAL and/or CAR T are generated by fusing different leucine zippers to the hinge region of the human CD8a chain and transmembrane and cytoplasmic regions of the human CD28, or CD3z signaling endo-domains. All CAL and/or CARs can contain a myc tag to verify surface expression. Besides, these primary T cells will also be fused to mCherry after CD3z chain to visualize expression.

zipFv Construct Design: The zipFv molecules contain an scFv against CD127 fused to an SYN2 (to stimulate the CD3z SYN1 zipCAR), and an scFv against CD45RO fused to a JUN zipper (to stimulate the costimulatory FOS zipCAR). The scfvs sequences are available and can be constructed rapidly through commercial DNA synthesis²⁸.

zipCAR transduction: Human PBMC are purified using separation kits. are introduced into primary human T cells via retroviral transduction. Expression of zipCARs is quantified via myc and V5-tag immunostaining and flow cytometry. As controls, a zipCAR that contains both CD3z and CD28 (i.e. SYN1-CD28-CD3z) can be used. This zipCAR cannot perform logic computation. A CD45RO-SYN2 or CD127-SYN2 zipFv can be added to ensure that zipCARs can kill CD45RO+ or CD127+ cells. These controls will also serve as reference for specificity.

Evaluate the Activity and Efficacy of Anti-CD127+/CD45RO+CAR T Cells In Vitro

Co-Culturing PBMCs with CD127+/CD45RO+ Cells with SUPRA CAR T Cells to Investigate the Specificity of Targeting

This experiment examines whether this system specifically kills CD127+ and CD45RO+ double positive T cells amongst a large population of PBMCs. PBMCs are obtained either from known VCA recipients or Mass General Blood Bank Center after being approved by IRB. First, human CD45RO+ and CD127+ are stained to identify the initial percentage of memory cells in the whole population (CD45RO/CD127 double positive vs double neg). Next, PBMCs are co-cultured with zipCAR expressing CD8+ T cells with zipFvs (anti CD127 and anti CD45RO) with the same condition. At the study time points (24, 48, 72 hrs) significant changes in the ratio of CD45RO/CD127+/+ to −/− will be considered as an indicator of whether SUPRA CARs were efficient in targeting specific T cells rather than non-specific cytotoxic effects. Additionally, engineered T cells and zipFvs will be cocultured with the memory T cells at 3 different zipFvs concentrations (5, 25, 50 ng/well) to determine the correlation between cell killing and zipFv concentration. T cell activation (CD69 expression, IL-2 and IFN-g in the media) and cytotoxicity against memory T cells (no. of remaining live cells) will also be measured. The proliferation of T cells will be measured by cell counting. All conditions will be tested at least in triplicate.

TABLE 1 outline of experiments 1 PBMC - Ctrl 2 PBMC + SUPRA CAR T 3 PBMC + SUPRA CAR T + scFv against CD45RO 4 PBMC + SUPRA CAR T + scFv against CD127 5 PBMC + SUPRA CAR T + scFvs CD127 + CD45RO

To Evaluate and Characterize the Effect of Memory T Cells on the HSCT Engraftment.

Generation of a Humanized Mouse Model with the VCA-Recipients PBMCs

To examine the effect of CD45RO/CD127 targeting with CAR T cells on the graft take and mixed chimerism sustainability, a humanized mouse model is generated by utilizing the PMBCs of VCA-recipients (FIGS. 8A, 8B). The humanized PBMC model has the fastest engraftment rate using adult PBMCs and enables studies that require a strong memory T cell function. VCA recipients' PBMCs hold an alloreactive population of memory T cells (CD45RO+/CD127+) and makes this a reliable model for studying their depletion effects on the rejection of the graft. The number of mice used per group in this study is chosen based on the recommendations from previous studies²⁹. In brief, irradiated NSG (MHC I/II K/O) (3 Gy) mice will be injected with 5×105 PBMC cells and human peripheral reconstitution is measured by flow cytometry every other week³⁰.

Testing the effects of memory T cells depletion on mixed chimerism establishment cells in a skin graft humanized mouse model: The NSG (MHC I/II K/O) mouse model that was generated by using the VCA-recipient PBMCs are used as the foundation for induction of mixed chimerism. The presence of memory T cells in the human PBMCs of this model imitates the challenges for induction of mixed chimerism in human subjects. NSG Hu-PBMCs are treated with/without (ctrl) anti CD45RO/CD127 CAR T to deplete the memory compartment of their T cells population (FIGS. 8A, 8B).

Fetal CD34+ HSC (and skin tissue) can be obtained from Advanced Bioscience Resources. Fetal HSCs are utilized to evaluate the generation and establishment of multi-lineage engraftment by using previously published Mixed Chimerism Induction Protocols (MCIP) in mice 23. In short, male mice are treated with anti CD154, anti-CD40, CTLA-4 and receive 3 Gy TBI. Six hours later they receive 2×105CD34+ HSC bone marrow cells. Engraftment is determined by measuring the presence of donor myeloid and lymphoid cells at different time points. Mixed chimerism is defined as at least 5% of leukocytes in each of the lineages being donor derived. Less than 5% or loss of multi-lineage status is considered as failure of chimerism.

Next, the aforementioned humanized mouse model (VCA-Recipient PBMCs-CD34 HSC) is utilized as a skin graft model to test the effect of depleting CD45RO/CD127 cells on allograft survival. Fetal skin graft is obtained from Advanced Bioscience Resources from the same CD34+ HSC donor. At week 10 post PBMC injection, skin grafts from the HSC donor and another allogeneic source are used to test whether nonspecific depletion of memory T cell repertoire affects the skin grafts survival (FIG. 8, 9). Mice are followed until graft rejection or POD 100.

Skin grafting will be conducted using previously published protocols³¹. In short, 2 pieces of tail skin from the donor are transplanted on the back of one recipient. Following dressing removal (POD7), mice and their skin grafts are evaluated daily. Graft failure is determined by the physical appearance of the graft; including color, texture, and contour. Histology is also performed to provide supportive data regarding the clinical signs of graft rejection or engraftment. Grafts with more than 50% shrinkage in size or change in color and flexibility (dark and firm grafts) are considered as failed. Skin graft are followed for 100 days.

TABLE 2 outline of experiments Skin # N Recipient (NSG-Hu-VCA-Rec.-PBMCs) Mixed Chimerism Grafts 1 8 Control - No treatment (MCIP) + 2 8 Control - zipFv (anti CD45RO/ PBMCs + HSCT + (MCIP) + CD127) 3 8 Control - SUPRA CAR T PBMCs + HSCT + (MCIP) + 4 8 SUPRA CAR T + zipFv (anti PBMCs + HSCT + (MCIP) + CD45RO/CD127) 5 8 SUPRA CAR T + zipFv (anti No PBMCs + HSCT, No MCIP + CD45RO/CD127) 6 8 SUPRA CAR T + zipFv (anti No MCIP + CD45RO/CD127) 7 8 SUPRA CAR T + zipFv (anti ⅓ of regular dose MCIP + CD45RO/CD127)

Off-target toxicity occurs when T cells are not able to distinguish normal cells from target cells. In this case CD45RO is the marker that is mainly expressed on the T cells but CD127 is a more common surface marker and can be found on B and dendritic cells. Even though the “Combinatorial AND Logic” CAR system that is described herein has shown to be highly specific, there is a chance for off-target effects in this system. As an alternative approach, all of the CD45RO+ population can be targeted, which consists of not only central memory T cells but also targets effector T cells as well. This will cause more T cells to be targeted nonspecifically but simultaneously might decrease the chance of immune reaction against the graft.

Some of the CAR T effector cells will express memory phenotype such as CD45RO/CD127 after remaining activated for a period of time. This means that CAR T cells might attack each other after changing phenotype. Long activation of CAR T cells within the patient's body has always been a matter of concern. This hypothetical situation can be considered as a safety step as CAR T cell will clear out themselves after completing their task which is destroying memory T cells. On the other hand, if turns out to be true, this self-eliminatory phenomenon might increase the number of CAR T cells that are needed to effectively treat the patients.

Benchmark for success: Blood drawn from the vain tail (40 ul) is used every other week for the assessment of peripheral blood chimerism. Monoclonal anti-donor HLA antibody is used for monitoring of chimerism in the various leukocyte lineages, including CD3+/CD8+, CD3+/CD4+, CD3+/CD8−/CD4− (naive T cells), T-Regs, naive and mature B cells (CD19+/CD78− and CD19−/CD78+). Animals are considered chimeric when at least 5% of WBCs in all of these lineages are donor derived. Additionally, the systemic immune status of humanized mice is assessed pre-HSC transplantation and at week 4 and week 8 post-transplant by carboxyfluorescein diacetate succinimidyl ester (CFSE) mixed lymphocyte reaction (MLR) proliferation assays. MLR results are compared before PMBC-HSC transplantation and after study end point.

Determine the Efficacy of pMHC-CAR T-Effector Cells on Specific Depletion of Recipient Alloreactive T Cells and Establishment of Mixed Chimerism.

Described herein is a flexible CAL and/or CAR design that can target any alloantigen specific T cells. Due to the diversity of alloantigen-specific T cells in the context of transplantation, the CAL and/or CAR system that has the flexibility to locate and attack different alloreactive T cells simultaneously. CAL and/or CAR T-eff. cells with (donor) pMHC can target alloreactive T-cells.

The Effect of CAL and/or CAR T Cell Therapy on Targeting a Double Antigen MHC System (OTi and OTii)

In short, pMHC multimer attached to adaptor molecule (e.g., the CAL)+CAR T effector cell is used in the single and double specific antigen-MHC systems. The antigen-specific CAL and/or CAR system can be tested in vitro and in the OTi & ii transgenic mouse model. This model is genetically engineered such that all T cells express receptors that are specific for recognizing chicken ovalbumin peptides (257-264 (OTi) and 329-337 (OTii) in the context of H2Kb (OTi) and I-A b (OTii)). The T cells of these mice models are engineered to express TCRs that specifically recognize pMHC class I/II+ Ovalbumin peptides. More importantly, this highly controlled model can be leveraged establish correlation between T cell activities and CAL and/or CAR T cell experimental parameters, such coculture conditions, pMHC affinity, pMHC multimer concentration.

Design, build, and test anti-FITC CAR/pMHC (e.g., the CAL) system in vitro against antigen specific T cells.

As mentioned above, older generations of CAR T cells needed to be genetically redesigned to be able to recognize new or multiple targets. To overcome this challenge, mouse split universal CD8 CAR is generated as described herein. For generating mouse FITC-CAR similar steps that were described previously for human anti-FITC CAR can be utilized²⁰. To evaluate the efficacy of C67BL/6 mouse anti-FITC CAR T cells efficacy, the experiments that are conducted and mentioned in the section above with human CAR T cells (FIG. 1-5) are repeated with mouse CAR T cells.

TABLE 3 outline of experiments Group N Recipient (B6-wild type + OTi/ii) 1 6 Control - No CAR/pMHC 2 6 Control + FITC pMHC multimer 3 6 Control + CAR T (no multimer) 4 6 CAR T + FITC pMHC multimer)

Evaluate the efficacy of CAR T-eff.+ FITC-pMHC tetramers (e.g. a CAL) for targeting alloreactive T-cells in the OTi and OTii transgenic mice.

Ovalbumin-pMHC-CAR T-cells effective in recognizing and destroying OTi and OTii T cells a mixture of the OTi and OTii T-cells are transferred to the wild type mice (B6) recipients. The read out of this experiment is the cell count of OTi & ii before and after CAL and/or CAR T cell therapy. The results show the effect of the CAR T-eff cells with the Ovalbumin peptide+MHC tetramer (e.g., the CAL) is successful in targeting the specific clone of OTi & ii cells.

Establish the potency of multiple peptide/multiple MHC (e.g., CAL)-CAR T cell therapy for targeting alloreactive T cell repertoire on mixed chimerism and skin graft survival.

The most significant barrier against graft take is full mismatch across MHCs of donors and recipients³². This challenge is addressed herein by using MHC (e.g., CAL)+CAR T cells to target alloreactive T cells. These MHC multimers are commercially available and can be readily conjugated with FITC labeling to be recognized by the anti FITC CAR T cells. Alternatively, there are many other available options for split universal CAL and/or CAR technology including leucine zippers which provide the option of tuning the affinity of the binding between adaptor (e.g., CAL) and CAR. On the other hand, Minor Histocompatibility Antigens (MiHA) are short immunogenic peptides originating from digested intra/extracellular proteins presented by Major Histocompatibility Complex (human leukocyte antigen). Disparities in minor histocompatibility antigens between individuals who are even MHC matched can induce an immune response after transplantation. These differences have shown to impede tolerance induction through mixed chimerism 33.

Generating a donor-recipient disparities peptide library for evaluating the efficacy of targeting alloreactive T cells in a humanized mouse model.

The role of the default peptides on these exchangeable-peptide MHC systems can be switched with the desired peptides (from the donor) using UV light^(34,35). The large collection of peptides associated with human leukocyte antigens (HLA) is referred to as the human immunopeptidome. The desired peptides for this study will are generated by the previously established protocols for immune-peptidome library generation^(36,37,38).

Generating tissue-specific immune peptide library from the donor and recipient.

Utilizing previously established protocols we can extract and identify the immunopeptidome of the donor and the recipient. We then identify the peptide sequences that are distinctively different between the donor and the recipient using software tools (below). These disparities in the donors' and the recipients' immune-peptide sequences have the potential to mediate immune rejection. The sequences of the peptides from the donor-recipient disparity peptide library are generated in practical scale by commercially available services³⁹ and loaded on peptide-exchangeable MHC multimers (e.g., a CAL) (QuickSwitch MBL international or Biolegend FlexTetramer) to ultimately be bound to CAR T cells and used for targeting alloreactive T cells in the aforementioned in vitro and in vivo models (FIG. 8, 9). In brief, donor and recipient MHCs associated peptides are isolated independently by immunoaffinity purification using the anti-HLA monoclonal antibodies³⁶. Eluted peptides are identified by different LC-MS/MS systems in DIA (Data Independent A) mode. Mass Spectrometry output files are converted, searched, and statistically validated using software tools (NETMHC, SysteMHC). The identified peptides are then clustered (by GibbsCluster v.1) and annotated by length and predicted MHC binding affinity (NetMHC v.4). The final list of high-confidence donor/recipient MHC-associated peptides is compared between the recipient and the donor and the disparate sequences used for building high-quality donor-recipient disparity peptide library, which is employed as a source for generating peptides to be combined with MBL QuickSwitch (or Bio-legend FlexT) MHC tetramer system.

TABLE 4 outline of experiments Skin Group N Recipient (NSG-Hu-VCA-Rec.-PBMCs) Mixed Chimerism Grafts 1 8 Control - No treatment (MCIP) + 2 8 SUPRA CAR T + FITC/(peptide MHC PBMCs + HSCT + (MCIP) + multimer 3 8 Control - SUPRA CAR T PBMCs + HSCT + (MCIP) + 4 8 SUPRA CAR T + FITC/(peptide MHC PBMCs + HSCT + (MCIP) + multimer) 5 8 SUPRA CAR T + FITC/(peptide MHC No PBMCs + HSCT, No MCIP + multimer) 6 8 SUPRA CAR T + FITC/(peptide MHC No MCIP + multimer) 7 8 SUPRA CAR T + FITC/(peptide MHC ⅓ of regular dose MCIP + multimer)

Evaluating the efficacy of targeting alloreactive T cells in the multiple antigen/MHC setting on mixed chimerism establishment in a skin graft humanized mouse model.

In the actual context of allotransplantation, both MHC mismatch and MiHA will be a potential source of immune response and rejection. This experiment addresses the question of whether combing the donor-recipient disparate peptide sequences with exchangeable-peptide MHC system (e.g., a CAL) can be used in combination with split CAR T cells to target alloreactive T cells.

Prior to initiating this experiment, human fetal HSC+skin graft from the same donor is obtained from Advanced Bioscience Resources (CA, US). HLA typing of the donor cells is done and used for generating donor (FITC+exchangeable-peptide) MHC multimer. Peptides that are used with these multimer are generated based on the results of the work described above herein. Briefly, the exchangeable-peptide MHC (e.g., the CAL) is exposed to UV for the dissociation of their default UV-labile peptide. The disparity peptide library provides the desired peptides to replace the default peptide of this system. Using the same double humanized PBMC/HSCT mouse model the efficacy of this donor/recipient disparity peptide library+MHC is evaluated for specific targeting of alloreactive memory T cells and skin allograft survival. As shown in FIGS. 8A-8B, Hu-PBMC treated with exchanged peptide MHC (e.g., CAL)+CAR T. Subsequently, HSCT with fetal HSC is performed and 6 weeks later skin from the same HSC donor is grafted on the back of the mice. Presence of mixed chimerism and graft survival is evaluated using the criteria that was mentioned earlier herein.

The immunopeptidome library is novel and has not previously been adapted for the context of transplantation. As an alternative to immunopeptidome mapping and disparity library generation, we can apply whole-peptidome (donor-recipient) disparity library, a well-known method.

Specificity and killing efficiency of CAL and/or CAR T cells, and chimerism levels can be studied in vitro and in vivo. Peripheral mechanisms of rejection or regulation can be assessed with serial mixed lymphocyte reaction (MLR) assays, at experimental endpoint. For our in vivo studies, our standard clinical definition of tolerance in rodents, which will serve as a benchmark, is rejection and immunosuppression-free survival for 100 days post-HSCT will serve as a benchmark. Skin grafts are assessed as mentioned earlier, based on the color, texture and size of shrinkage.

Impact: Described herein is a novel adaptive immune cell therapy strategy, making use of the CAL and/or CAR T-cell technology, for VCA tolerance induction. Musculoskeletal extremity injuries, including traumatic amputations are frequently seen in the pediatric burn and trauma population. These patients often struggle with severe functional and psychosocial challenges and decreased quality and quantity of life. In turn, the bottleneck to wide application of VCA, with its obvious benefits, would be lifted and effect a paradigm shift in the current management of devastating pediatric burns, which often result in severe disfigurement and amputations. Additionally, successful results enable VCA to be performed for trauma or congenital defects.

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Example 9

Jurkat+pMHC-OTi

Described herein is the investigation of whether killer CAR T cells activation by binding to the adaptor (pMHC, e.g, a CAL))+ Target. The experimental design is depicted in FIG. 11.

FITC-conjugated tetramer mediated activation was verified (FIG. 12) and followed in a time course (FIG. 13). Anti-FITC Jurkat vs OT-I experiments were conducted using 100 k cells anti-FITC CAR Jurkat, 100 k cells splenocytes from OT-I mouse, and 5 μg/mL to 78 ng/mL FITC-conjugated tetramer (positive, H2Kb-SIINFEKL (SEQ ID NO: 2750); negative, I-Ab-AAHAEINEA (SEQ ID NO: 2751)). Total Jurkat cell counts at 24 hours (FIG. 14) and tetramer staining levels (FIG. 15) were also determined. The levels of tetramer and CD69 were also determined (FIG. 16).

This experiment demonstrated that Jurkat cells get activated in a dose dependent fashion up to 1 μg/ml of tetramer and OTi binding and staining is dose dependent. No cytotoxicity effects were seen on the killer cells and no prominent activation was seen on the target cells.

Primary T Cells-OTi

Experimental design is shown in FIG. 17. Human CD8 pMHC-CAR T killing is highly specific and MHC-CAR T cell does not kill bystander CD4 T cells (FIG. 18). No cytotoxicity effect was seen on human CD8 pMHC-CAR T (FIG. 19). Cytotoxicity against CAR T cells was long-lived (FIG. 20) and levels of CD69 on OTi CD8 T cells was followed (FIG. 21).

This experiment demonstrated activation of target cells with no significant cytotoxicity on the killer cells. It is possible that Killer Cells activation might be suboptimal under these conditions and use of primary T cells could include fresh, syngeneic cells for improved activation.

Example 10

1E6 T cell clone is a well-established CD8+ specific for the major β-cell autoantigen, preproinsulin (PPI). The 1E6 clone mediates β-cell-specific killing via recognition of a highly distinctive HLA A*0201-presented signal peptide epitope (PPI15-24) that exhibits glucose-dependent presentation on the surface of human β-cells (9). The inventors generated and tested a split pMHC-CAR that targets 1E6 T-cells through A*0201+peptides. Effective killing of 1E6 T-cells was observed (FIG. 29)

Example 11

CALs comprising ovalbumin MiHA (against OTi or OTii) were constructed (FIG. 26). pMHC tetramer+FITC (adaptor) binds to the target cells (OTi) in a dose dependent fashion, while OTii specific tetramer does not (FIG. 27). Binding of Jurkat+pMHC to OTi cells does not change Jurkat live count (FIGS. 28B and 30). However, human CD8 CAR T were cytotoxic with high specificity (FIGS. 31 and 32). Cytotoxicity was not seen with ctrl tetramer. No cytotoxicity was seen on killer CAR T cells or bystander CD4 T Cells after co-culturing with pMHC and splenocytes. 

1. A composition, comprising: a. a TCR recognition domain; and one or both of: b. an intracellular signaling domain; and c. a biomolecular interaction domain.
 2. The composition of claim 1, comprising a TCR recognition domain and a biomolecular interaction domain.
 3. The composition of claim 1, comprising a TCR recognition domain and an intracellular signaling domain.
 4. The composition of claim 1, wherein the biomolecular interaction domain of c) is a first-type biomolecular interaction domain and the composition further comprises a signaling polypeptide comprising a second-type biomolecular interaction domain and an intracellular signaling domain; wherein the first-type and second-type biomolecular interaction domains bind specifically to each other.
 5. The composition of claim 1, wherein the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MHC-peptide complex; a featureless peptide MHC; or a MHC-peptide fusion.
 6. The composition of claim 1, wherein the TCR recognition domain comprises a CD1 domain or a CD1 domain-ligand complex or fusion.
 7. The composition of claim 6, wherein the CD1 is CD1d.
 8. The composition of claim 1, wherein the TCR recognition domain is a monomer, dimer, trimer, tetramer, pentamer, dextramer or other oligomer form.
 9. The composition of claim 1, a. wherein the biomolecular interaction domain(s) is a leucine zipper, or any binding pair of biomolecular interaction domains are collectively a pair of leucine zippers; b. wherein the biomolecular interaction domain(s) is a BZip (RR) and/or a AZip (EE), or any binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE); c. wherein the biomolecular interaction domain(s) is a PSD95-D1g1-zo-1 (PDZ) domain; d. wherein the biomolecular interaction domain(s) is a streptavidin and/or a streptavidin binding protein (SBP) or any binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP); e. wherein the biomolecular interaction domain(s) is a FKBP-binding domain of mTOR (FRB) and/or a FK506 binding protein (FKBP) or any binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding protein (FKBP); f. wherein the biomolecular interaction domain(s) is a cyclophilin-Fas fusion protein (CyP-Fas) and/or a FK506 binding protein (FKBP) or any binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion protein (CyP-Fas) and a FK506 binding protein (FKBP); g. wherein the biomolecular interaction domain(s) is a calcineurin A (CNA) and/or a FK506 binding protein (FKBP) or any binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding protein (FKBP); h. wherein the biomolecular interaction domain(s) is a gibberellin insensitive (GIA) and/or a gibberellin insensitive dwarf1 (GID1) or any binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1); i. wherein the biomolecular interaction domain(s) is a Snap-tag and/or a Halo tag, or any binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag; j. wherein the biomolecular interaction domain(s) is a T14-3-3-cdeltaC and/or a C-Terminal peptides of PMA2 (CT52), or any binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52); k. wherein the biomolecular interaction domain(s) is a PYL and/or a ABI, or any binding pair of biomolecular interaction domains are collectively a PYL and a ABI; l. wherein the biomolecular interaction domain(s) is a nucleotide tag and/or a zinc finger domain, or any binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain; m. wherein the biomolecular interaction domain(s) is a nucleotide tag, or any binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags; n. wherein the biomolecular interaction domain(s) is a Fluorescein isothiocyanate (FITC) and/or a FITC binding protein or any binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or o. wherein the protein interaction domain(s) is a (R)-Phycoerythrin (R-PE/PE) and/or a R-PE/PE binding protein or any binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE binding protein.
 10. The composition of claim 9, wherein the nucleotide tag is a DNA tag or dsDNA tag.
 11. The composition of claim 2, further comprising a cell expressing or comprising the signaling polypeptide.
 12. The composition of claim 11, wherein the TCR recognition domain is allogeneic, autologous, or xenogenic to the cell.
 13. The composition of claim 11, wherein the TCR recognition domain is synthetic.
 14. The composition of claim 11, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogenic to the cell.
 15. The composition of claim 11, wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.
 16. The composition of claim 11, wherein the cell is a NK cell, dendritic cell, regulatory T cell, or effector T cell.
 17. The composition of claim 11, wherein the cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.
 18. A method of treating or preventing an autoimmune disease or condition; T cell mediated inflammation or immune response; malignant T cell condition; transplant rejection; or GvHD in a subject in need thereof, the method comprising administering to the subject a composition of claim
 1. 19. The method of claim 18, wherein the transplant is an allogeneic hematopoietic stem cell or solid organ transplantation.
 20. The method of claim 18, wherein the malignant T cell condition is T cell acute lymphoblastic leukemia or T cell lymphoblastic lymphoma.
 21. The method of claim 18, wherein the autoimmune disease is type 1 diabetes, vitiligo, multiple sclerosis, alopecia, celiac disease, pemphigus, rheumatoid arthritis, or scleroderma.
 22. The method of claim 18, wherein the T cell mediated immune response is an anti-drug specific response to a biologic, cell therapy, and/or gene therapy.
 23. The method of claim 22, wherein the biologic, cell-therapy, or gene therapy is an adeno-associated virus (AAV) gene therapy, a genome editing agent, or enzyme replacement therapy.
 24. The method claim 18, wherein the disease is type 1 diabetes and the TCR recognition domain comprises sequences with at least 95% sequence identity to: one or more of SEQ ID NOs: 8-17; HLA-A*0201 and at least one of SEQ ID NOs: 2013-2016 and 2031-2033; or HLA-A*02:01 and at least one of SEQ ID NOs: 20128-2129.
 25. The method of claim 18, wherein the disease is vitiligo and the TCR recognition domain comprises sequences with at least 95% sequence identity to: SEQ ID NO: 18, 19, and one of 20-22; or HLA-A*0201 and SEQ ID NO: 2018; or HLA-A*0301 and SEQ ID NO: 2019; or comprises HLA-A*2402 and SEQ ID NO: 2020; or HLA-A*0101 and SEQ ID NO:
 2021. 26. The method of claim 18, wherein the method is a method of treating and/or preventing GvHD and the TCR recognition domain comprises sequences with at least 95% sequence identity to: HLA-A*0101 and at least one of SEQ ID NOs: 2034-2037; or HLA-B*0702 and SEQ ID NO: 2038; or HLA-B*0801 and SEQ ID NO:
 2039. 